TY  - GEN
AU  - Ćirković-Veličković, Tanja
AU  - Gnjatović, Marija
AU  - Ćujić, Danica
AU  - Todorović, Aleksandra
AU  - Stanić-Vučinić, Dragana
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Simović, Ana
AU  - Radosavljević, Jelena
AU  - Aćimović, Jelena M.
AU  - Radomirović, Mirjana Ž.
AU  - Stojanović, Marijana
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/860
AB  - Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.
T1  - Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus
UR  - https://hdl.handle.net/21.15107/rcub_intor_860
ER  - 

@misc{
author = "Ćirković-Veličković, Tanja and Gnjatović, Marija and Ćujić, Danica and Todorović, Aleksandra and Stanić-Vučinić, Dragana and Đukić, Teodora and Mladenović, Maja and Vasović, Tamara and Stojadinović, Marija and Krstić-Ristivojević, Maja and Jovanović, Vesna and Simović, Ana and Radosavljević, Jelena and Aćimović, Jelena M. and Radomirović, Mirjana Ž. and Stojanović, Marijana",
year = "2023",
abstract = "Novi korona virus (SARS CoV-2) koji se pojavio u Vuhanu 2019. godine pripada grupi jednolančanih RNK virusa [1]. Predstavlja novi infektivni agens za humanu populaciju i veoma je brzo detektovan u velikom broju zemalja. Uzročnik je respiratornih infekcija koje mogu da budu praćene i veoma teškom kliničkom slikom. Brzo širenje, odsustvo imuniteta na ovaj virus i odsustvo pouzdanih testova za detekciju virusa u trenutku izbijanja pandemije su bolest izazvanu ovim virusom brzo pretvorili u zdravstveni i društveni problem najvišeg prioriteta na globalnom nivou. Iako su najveće biotehnološke kompanije ubrzano počele sa razvojem i masovnom proizvodnjom dijagnostičkih testova i vakcina, njihova dostupnost u trenucima najveće potražnje je i dalje nedovoljna, a cene istih su limitirajući faktor za bolju kontrolu bolesti i širenja pandemije [2]. Razvoj sopstvenih i održiva proizvodnja testova i vakcina za COVID-19 su od velikog društvenog značaja. Važan preduslov za održivu proizvodnju testova je dostupnost rekombinantnih antigena virusa i mogućnost proizvodnje istih na velikoj skali za potrebe proizvodnje domaćih testova. Ovim tehničkim rešenjem se opisuje dobijanje dva ključna antigena novog korona virusa rekombinantnom tehnologijom i njihova primena u serološkom ELISA testu koji proizvodi Institut za primenu nuklearne energije, INEP, kao i za dobijanje reagenasa za detekciju antigena novog korona virusa (specifičnih antitela). U prvoj fazi, optimizovane su sekvence proteina koje su podigle osetljivost postojećih seroloških testova. Inovativnost našeg pristupa se ogleda i u razrađenim eksperimentalnim protokolima za dobijanje rekombinantnih proteina nukleokapsida na velikoj skali, kao i u solubilnoj formi, što olakšava postupak prečišćavanja. Izbor fragmenta nukleokapsida koji se heterologo eksprimira u solubilnoj formi, a specifično detektuje antitela i generiše jak imuni odgovor tokom imunizacije životinja (imunogenost) na osnovu pregleda poznatih epitopskih sekvenci je ključna inovacija ovog tehničkog rešenja. Ovo je prvi primer uspešno primenjenog rekombinatnog proteina proizvedenog u Srbiji u dijagnostičkom testu koji je registrovankod Agencije za lekove i medicinska sredstva Republike Srbije (broj rešenja 515-02-02370-21-002), a koji je primenu našao i na međunarodnom nivou.",
title = "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus",
url = "https://hdl.handle.net/21.15107/rcub_intor_860"
}

Ćirković-Veličković, T., Gnjatović, M., Ćujić, D., Todorović, A., Stanić-Vučinić, D., Đukić, T., Mladenović, M., Vasović, T., Stojadinović, M., Krstić-Ristivojević, M., Jovanović, V., Simović, A., Radosavljević, J., Aćimović, J. M., Radomirović, M. Ž.,& Stojanović, M.. (2023). Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. .
https://hdl.handle.net/21.15107/rcub_intor_860

Ćirković-Veličković T, Gnjatović M, Ćujić D, Todorović A, Stanić-Vučinić D, Đukić T, Mladenović M, Vasović T, Stojadinović M, Krstić-Ristivojević M, Jovanović V, Simović A, Radosavljević J, Aćimović JM, Radomirović MŽ, Stojanović M. Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. 2023;.
https://hdl.handle.net/21.15107/rcub_intor_860 .

Ćirković-Veličković, Tanja, Gnjatović, Marija, Ćujić, Danica, Todorović, Aleksandra, Stanić-Vučinić, Dragana, Đukić, Teodora, Mladenović, Maja, Vasović, Tamara, Stojadinović, Marija, Krstić-Ristivojević, Maja, Jovanović, Vesna, Simović, Ana, Radosavljević, Jelena, Aćimović, Jelena M., Radomirović, Mirjana Ž., Stojanović, Marijana, "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus" (2023),
https://hdl.handle.net/21.15107/rcub_intor_860 .

TY  - JOUR
AU  - Khulal, Urmila
AU  - Stojadinović, Marija M.
AU  - Prodić, Ivana
AU  - Rajković, Andrea
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/766
AB  - The digestion stability of allergen pairs, tropomyosin, TM (fish and seafood allergen), and myosin light chain, MLC (chicken meat allergen) is compared among vertebrates and invertebrates in raw and cooked food matrix under standardized simulated in vitro gastrointestinal (GI) digestion. SDS-PAGE followed by multiple TM and MLC-specific antibodies in semidry WB revealed pepsin resistance of invertebrate TMs (abalone, oyster, shrimp) under diet-relevant conditions (raw, cooked). Vertebrate TMs (chicken, pork, beef) were less stable to digestion except that the raw chicken TM was observed pepsin resistant (not diet-relevant). Vertebrate (chicken) MLC was thermally stable. A new 28 kDa protein bound to anti-MLC antibody in cooked chicken and pork; could be the aggregates of MLC. Raw shrimp MLC showed pepsin resistance among invertebrates. A good correlation was observed between combined resistance of TM and MLC to GI digestion following the diet-relevant thermal treatment and reported protein allergenicity among vertebrates and invertebrates.
PB  - Elsevier
T2  - Food Chemistry
T1  - Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix
SP  - 134981
VL  - 405
DO  - 10.1016/j.foodchem.2022.134981
ER  - 

@article{
author = "Khulal, Urmila and Stojadinović, Marija M. and Prodić, Ivana and Rajković, Andrea and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "The digestion stability of allergen pairs, tropomyosin, TM (fish and seafood allergen), and myosin light chain, MLC (chicken meat allergen) is compared among vertebrates and invertebrates in raw and cooked food matrix under standardized simulated in vitro gastrointestinal (GI) digestion. SDS-PAGE followed by multiple TM and MLC-specific antibodies in semidry WB revealed pepsin resistance of invertebrate TMs (abalone, oyster, shrimp) under diet-relevant conditions (raw, cooked). Vertebrate TMs (chicken, pork, beef) were less stable to digestion except that the raw chicken TM was observed pepsin resistant (not diet-relevant). Vertebrate (chicken) MLC was thermally stable. A new 28 kDa protein bound to anti-MLC antibody in cooked chicken and pork; could be the aggregates of MLC. Raw shrimp MLC showed pepsin resistance among invertebrates. A good correlation was observed between combined resistance of TM and MLC to GI digestion following the diet-relevant thermal treatment and reported protein allergenicity among vertebrates and invertebrates.",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix",
pages = "134981",
volume = "405",
doi = "10.1016/j.foodchem.2022.134981"
}

Khulal, U., Stojadinović, M. M., Prodić, I., Rajković, A.,& Ćirković-Veličković, T.. (2023). Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix. in Food Chemistry
Elsevier., 405, 134981.
https://doi.org/10.1016/j.foodchem.2022.134981

Khulal U, Stojadinović MM, Prodić I, Rajković A, Ćirković-Veličković T. Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix. in Food Chemistry. 2023;405:134981.
doi:10.1016/j.foodchem.2022.134981 .

Khulal, Urmila, Stojadinović, Marija M., Prodić, Ivana, Rajković, Andrea, Ćirković-Veličković, Tanja, "Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix" in Food Chemistry, 405 (2023):134981,
https://doi.org/10.1016/j.foodchem.2022.134981 . .

TY  - JOUR
AU  - Khulal, Urmila
AU  - Stojadinović, Marija M.
AU  - Prodić, Ivana
AU  - Rajković, Andrea
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/767
AB  - The digestion stability of allergen pairs, tropomyosin, TM (fish and seafood allergen), and myosin light chain, MLC (chicken meat allergen) is compared among vertebrates and invertebrates in raw and cooked food matrix under standardized simulated in vitro gastrointestinal (GI) digestion. SDS-PAGE followed by multiple TM and MLC-specific antibodies in semidry WB revealed pepsin resistance of invertebrate TMs (abalone, oyster, shrimp) under diet-relevant conditions (raw, cooked). Vertebrate TMs (chicken, pork, beef) were less stable to digestion except that the raw chicken TM was observed pepsin resistant (not diet-relevant). Vertebrate (chicken) MLC was thermally stable. A new 28 kDa protein bound to anti-MLC antibody in cooked chicken and pork; could be the aggregates of MLC. Raw shrimp MLC showed pepsin resistance among invertebrates. A good correlation was observed between combined resistance of TM and MLC to GI digestion following the diet-relevant thermal treatment and reported protein allergenicity among vertebrates and invertebrates.
PB  - Elsevier
T2  - Food Chemistry
T1  - Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix
SP  - 134981
VL  - 405
UR  - https://hdl.handle.net/21.15107/rcub_intor_767
ER  - 

@article{
author = "Khulal, Urmila and Stojadinović, Marija M. and Prodić, Ivana and Rajković, Andrea and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "The digestion stability of allergen pairs, tropomyosin, TM (fish and seafood allergen), and myosin light chain, MLC (chicken meat allergen) is compared among vertebrates and invertebrates in raw and cooked food matrix under standardized simulated in vitro gastrointestinal (GI) digestion. SDS-PAGE followed by multiple TM and MLC-specific antibodies in semidry WB revealed pepsin resistance of invertebrate TMs (abalone, oyster, shrimp) under diet-relevant conditions (raw, cooked). Vertebrate TMs (chicken, pork, beef) were less stable to digestion except that the raw chicken TM was observed pepsin resistant (not diet-relevant). Vertebrate (chicken) MLC was thermally stable. A new 28 kDa protein bound to anti-MLC antibody in cooked chicken and pork; could be the aggregates of MLC. Raw shrimp MLC showed pepsin resistance among invertebrates. A good correlation was observed between combined resistance of TM and MLC to GI digestion following the diet-relevant thermal treatment and reported protein allergenicity among vertebrates and invertebrates.",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix",
pages = "134981",
volume = "405",
url = "https://hdl.handle.net/21.15107/rcub_intor_767"
}

Khulal, U., Stojadinović, M. M., Prodić, I., Rajković, A.,& Ćirković-Veličković, T.. (2023). Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix. in Food Chemistry
Elsevier., 405, 134981.
https://hdl.handle.net/21.15107/rcub_intor_767

Khulal U, Stojadinović MM, Prodić I, Rajković A, Ćirković-Veličković T. Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix. in Food Chemistry. 2023;405:134981.
https://hdl.handle.net/21.15107/rcub_intor_767 .

Khulal, Urmila, Stojadinović, Marija M., Prodić, Ivana, Rajković, Andrea, Ćirković-Veličković, Tanja, "Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix" in Food Chemistry, 405 (2023):134981,
https://hdl.handle.net/21.15107/rcub_intor_767 .

TY  - CONF
AU  - Jovanović, Vesna B.
AU  - Smiljanić, Katarina
AU  - Lujić, Tamara
AU  - Đukić, Teodora
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/779
AB  - Edible insects are recommended as a future food because of many reasons. The nutritional value of edible insects is one of criteria for the selection of seven most promising species among which is Tenebrio molitor. The objective of this study was to examine the effects of different flour Tenebrio molitor. Twelve different extraction conditions were set up in which three parameters were combined: pH extraction solutions (6, 8 and 12.5), temperature (30 °C and 60 °C), and ultrasound (US). Shotgun proteomics of trypsin digests profiled protein isolates. The highest protein yield was in extractions at pH 12.5. The temperature elevation and US application significantly increased the yield of isolated proteins at pH 12.5 but their solubility at the pH 7.4 was lower compared to isolates at pH 6 and 8. 1D-SDS-PAGE showed marked differences in protein profiles on various extraction conditions, with highest number of the distinctive bands at pH 8 at 30 °C. Shotgun proteomics showed that extraction condition at pH 12.5, on 30 °C has the highest numbers of different proteins, however, among the top 20 abundant proteins are chitin-associated proteins, allergens and proteinases, while at pH 8 these proteins are not enriched. Highly basic extraction significantly contributes to protein hydrolysis while application of US contributes to the protein cross-linking and this effect is more prominent at high temperatures.
C3  - Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021
T1  - Effects of extraction conditions on proteins' profiles of Tenebrio molitor
EP  - 53
SP  - 53
UR  - https://hdl.handle.net/21.15107/rcub_intor_779
ER  - 

@conference{
author = "Jovanović, Vesna B. and Smiljanić, Katarina and Lujić, Tamara and Đukić, Teodora and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Edible insects are recommended as a future food because of many reasons. The nutritional value of edible insects is one of criteria for the selection of seven most promising species among which is Tenebrio molitor. The objective of this study was to examine the effects of different flour Tenebrio molitor. Twelve different extraction conditions were set up in which three parameters were combined: pH extraction solutions (6, 8 and 12.5), temperature (30 °C and 60 °C), and ultrasound (US). Shotgun proteomics of trypsin digests profiled protein isolates. The highest protein yield was in extractions at pH 12.5. The temperature elevation and US application significantly increased the yield of isolated proteins at pH 12.5 but their solubility at the pH 7.4 was lower compared to isolates at pH 6 and 8. 1D-SDS-PAGE showed marked differences in protein profiles on various extraction conditions, with highest number of the distinctive bands at pH 8 at 30 °C. Shotgun proteomics showed that extraction condition at pH 12.5, on 30 °C has the highest numbers of different proteins, however, among the top 20 abundant proteins are chitin-associated proteins, allergens and proteinases, while at pH 8 these proteins are not enriched. Highly basic extraction significantly contributes to protein hydrolysis while application of US contributes to the protein cross-linking and this effect is more prominent at high temperatures.",
journal = "Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021",
title = "Effects of extraction conditions on proteins' profiles of Tenebrio molitor",
pages = "53-53",
url = "https://hdl.handle.net/21.15107/rcub_intor_779"
}

Jovanović, V. B., Smiljanić, K., Lujić, T., Đukić, T.,& Ćirković-Veličković, T.. (2021). Effects of extraction conditions on proteins' profiles of Tenebrio molitor. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021, 53-53.
https://hdl.handle.net/21.15107/rcub_intor_779

Jovanović VB, Smiljanić K, Lujić T, Đukić T, Ćirković-Veličković T. Effects of extraction conditions on proteins' profiles of Tenebrio molitor. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021. 2021;:53-53.
https://hdl.handle.net/21.15107/rcub_intor_779 .

Jovanović, Vesna B., Smiljanić, Katarina, Lujić, Tamara, Đukić, Teodora, Ćirković-Veličković, Tanja, "Effects of extraction conditions on proteins' profiles of Tenebrio molitor" in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021 (2021):53-53,
https://hdl.handle.net/21.15107/rcub_intor_779 .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Jovanović, Vesna B.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/780
C3  - Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021
T1  - Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting
EP  - 71
SP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_intor_780
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana and Đukić, Teodora and Vasović, Tamara and Jovanović, Vesna B. and Ćirković-Veličković, Tanja",
year = "2021",
journal = "Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021",
title = "Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting",
pages = "71-71",
url = "https://hdl.handle.net/21.15107/rcub_intor_780"
}

Smiljanić, K., Prodić, I., Đukić, T., Vasović, T., Jovanović, V. B.,& Ćirković-Veličković, T.. (2021). Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021, 71-71.
https://hdl.handle.net/21.15107/rcub_intor_780

Smiljanić K, Prodić I, Đukić T, Vasović T, Jovanović VB, Ćirković-Veličković T. Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021. 2021;:71-71.
https://hdl.handle.net/21.15107/rcub_intor_780 .

Smiljanić, Katarina, Prodić, Ivana, Đukić, Teodora, Vasović, Tamara, Jovanović, Vesna B., Ćirković-Veličković, Tanja, "Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting" in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021 (2021):71-71,
https://hdl.handle.net/21.15107/rcub_intor_780 .

TY  - CONF
AU  - MIhilović, Jelena
AU  - Đukić, Teodora
AU  - Smiljanić, Katarina
AU  - Apostolović, Danijela
AU  - Liu, Shu-Hua
AU  - Epstein, Michelle M.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/769
AB  - Peanut allergy affects approximately up to 3 % of children and up to 2 % of the adult world population, causing reactions ranging from mild to severe. Major peanut allergens are well characterized but little is known about their post-translational modifications and even less is known about the influence of thermal treatment on their profile. Protein post-translational modification patterns may differ between raw and thermally treated peanuts, which could affect its functional properties, such as allergic potential. In this study we combined proteomic and immunological methods to characterize the modifications or proteoforms of four major peanut allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6 in raw and roasted peanut. Bottom-up high-resolution accurate mass spectrometry and a specialized proteomics software package to identify, map and compare modifications of major peanut allergens between differently treated peanut kernels. Modification-specific antibody western blot was used to confirm the presence of modifications on major allergens in both extracts. Twenty different post-translational modifications in four prominent peanut allergens (Ara h 1-3, 6) were identified, while twelve were quantitatively compared between raw and roasted peanuts by high-resolution mass spectrometry and a proprietary proteomics software. post-translational modification specific antibodies confirmed the presence of these modifications in western-blots of raw and roasted peanuts. This study initiates appreciation of modifications and thermal processing affecting food quality, and development of state-of-the-art methodology in the risk assessment of allergen contamination.
C3  - FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia
T1  - Comparative quantitative immunoproteomic study of raw and roasted peanut major allergen modifications
EP  - 27
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_intor_769
ER  - 

@conference{
author = "MIhilović, Jelena and Đukić, Teodora and Smiljanić, Katarina and Apostolović, Danijela and Liu, Shu-Hua and Epstein, Michelle M. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Peanut allergy affects approximately up to 3 % of children and up to 2 % of the adult world population, causing reactions ranging from mild to severe. Major peanut allergens are well characterized but little is known about their post-translational modifications and even less is known about the influence of thermal treatment on their profile. Protein post-translational modification patterns may differ between raw and thermally treated peanuts, which could affect its functional properties, such as allergic potential. In this study we combined proteomic and immunological methods to characterize the modifications or proteoforms of four major peanut allergens - Ara h 1, Ara h 2, Ara h 3 and Ara h 6 in raw and roasted peanut. Bottom-up high-resolution accurate mass spectrometry and a specialized proteomics software package to identify, map and compare modifications of major peanut allergens between differently treated peanut kernels. Modification-specific antibody western blot was used to confirm the presence of modifications on major allergens in both extracts. Twenty different post-translational modifications in four prominent peanut allergens (Ara h 1-3, 6) were identified, while twelve were quantitatively compared between raw and roasted peanuts by high-resolution mass spectrometry and a proprietary proteomics software. post-translational modification specific antibodies confirmed the presence of these modifications in western-blots of raw and roasted peanuts. This study initiates appreciation of modifications and thermal processing affecting food quality, and development of state-of-the-art methodology in the risk assessment of allergen contamination.",
journal = "FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia",
title = "Comparative quantitative immunoproteomic study of raw and roasted peanut major allergen modifications",
pages = "27-27",
url = "https://hdl.handle.net/21.15107/rcub_intor_769"
}

MIhilović, J., Đukić, T., Smiljanić, K., Apostolović, D., Liu, S., Epstein, M. M.,& Ćirković-Veličković, T.. (2021). Comparative quantitative immunoproteomic study of raw and roasted peanut major allergen modifications. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia, 27-27.
https://hdl.handle.net/21.15107/rcub_intor_769

MIhilović J, Đukić T, Smiljanić K, Apostolović D, Liu S, Epstein MM, Ćirković-Veličković T. Comparative quantitative immunoproteomic study of raw and roasted peanut major allergen modifications. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia. 2021;:27-27.
https://hdl.handle.net/21.15107/rcub_intor_769 .

MIhilović, Jelena, Đukić, Teodora, Smiljanić, Katarina, Apostolović, Danijela, Liu, Shu-Hua, Epstein, Michelle M., Ćirković-Veličković, Tanja, "Comparative quantitative immunoproteomic study of raw and roasted peanut major allergen modifications" in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia (2021):27-27,
https://hdl.handle.net/21.15107/rcub_intor_769 .

TY  - CONF
AU  - Prodić, Ivana
AU  - Smiljanić, Katarina
AU  - Hoffmann-Sommergruber, Karin
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/755
AB  - Background: In vitro pepsin digestion is important factor when assessing protein foodallergenicity. Roasted hazelnut is more common in human nutrition than a raw hazelnut;however, all studies were focused on Cor a 9 allergen obtained from a raw hazelnut. Thereare only two studies employing in vitro INFOGEST digestion harmonized protocol onhazelnut with its full matrix. The aim of this study was to assess immunoreactivity of rawand roasted hazelnut gastric digests and to compare secondary/tertiary structure of Cor a 9allergen purified from these two sources.Methods: Digestion resistant protein fragments were analysed by 1D/2D electrophoresis.Following digestion, IgE binding from patients’ pooled sera and by specific antibodies, wereassessed in ELISA and immunoblot. CD spectroscopy was applied for Cor a 9 structuralanalyses.Results: Cor a 11 and acidic forms of Cor a 9 were more prone to pepsin proteolysis, yettheir large fragments survived partially. Cor a 8 was protected by lipids, retaining capabilityto bind its specific antibody. Roasting did not significantly affect secondary structure of themost abundant hazelnut allergen, Cor a 9.Conclusion: Roasting of hazelnut seems to boost IgE binding derived from pooled sera ofhazelnut allergic patients with oral-gastric allergen digests.
PB  - INFOGEST Cost action, INRAE, Teagasc LTD.
C3  - Virtual International Conference on Food Digestion 6th and 7th May, 2021
T1  - Allergome of oral-gastric in vitro digest of roasted hazelnut shows stronger IgE binding compared to the raw counterpart
EP  - 62
SP  - 62
UR  - https://hdl.handle.net/21.15107/rcub_intor_755
ER  - 

@conference{
author = "Prodić, Ivana and Smiljanić, Katarina and Hoffmann-Sommergruber, Karin and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Background: In vitro pepsin digestion is important factor when assessing protein foodallergenicity. Roasted hazelnut is more common in human nutrition than a raw hazelnut;however, all studies were focused on Cor a 9 allergen obtained from a raw hazelnut. Thereare only two studies employing in vitro INFOGEST digestion harmonized protocol onhazelnut with its full matrix. The aim of this study was to assess immunoreactivity of rawand roasted hazelnut gastric digests and to compare secondary/tertiary structure of Cor a 9allergen purified from these two sources.Methods: Digestion resistant protein fragments were analysed by 1D/2D electrophoresis.Following digestion, IgE binding from patients’ pooled sera and by specific antibodies, wereassessed in ELISA and immunoblot. CD spectroscopy was applied for Cor a 9 structuralanalyses.Results: Cor a 11 and acidic forms of Cor a 9 were more prone to pepsin proteolysis, yettheir large fragments survived partially. Cor a 8 was protected by lipids, retaining capabilityto bind its specific antibody. Roasting did not significantly affect secondary structure of themost abundant hazelnut allergen, Cor a 9.Conclusion: Roasting of hazelnut seems to boost IgE binding derived from pooled sera ofhazelnut allergic patients with oral-gastric allergen digests.",
publisher = "INFOGEST Cost action, INRAE, Teagasc LTD.",
journal = "Virtual International Conference on Food Digestion 6th and 7th May, 2021",
title = "Allergome of oral-gastric in vitro digest of roasted hazelnut shows stronger IgE binding compared to the raw counterpart",
pages = "62-62",
url = "https://hdl.handle.net/21.15107/rcub_intor_755"
}

Prodić, I., Smiljanić, K., Hoffmann-Sommergruber, K.,& Ćirković-Veličković, T.. (2021). Allergome of oral-gastric in vitro digest of roasted hazelnut shows stronger IgE binding compared to the raw counterpart. in Virtual International Conference on Food Digestion 6th and 7th May, 2021
INFOGEST Cost action, INRAE, Teagasc LTD.., 62-62.
https://hdl.handle.net/21.15107/rcub_intor_755

Prodić I, Smiljanić K, Hoffmann-Sommergruber K, Ćirković-Veličković T. Allergome of oral-gastric in vitro digest of roasted hazelnut shows stronger IgE binding compared to the raw counterpart. in Virtual International Conference on Food Digestion 6th and 7th May, 2021. 2021;:62-62.
https://hdl.handle.net/21.15107/rcub_intor_755 .

Prodić, Ivana, Smiljanić, Katarina, Hoffmann-Sommergruber, Karin, Ćirković-Veličković, Tanja, "Allergome of oral-gastric in vitro digest of roasted hazelnut shows stronger IgE binding compared to the raw counterpart" in Virtual International Conference on Food Digestion 6th and 7th May, 2021 (2021):62-62,
https://hdl.handle.net/21.15107/rcub_intor_755 .

TY  - CONF
AU  - Prodić, Ivana
AU  - Smiljanić, Katarina
AU  - Nagl, Christoph
AU  - Hoffmann-Sommergruber, Karin
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/753
AB  - Cor a 8 is a relevant allergen that can cause severe allergic reactions. It is a 115 amino acid protein with a molecular mass of 9 kDa and is a member of the non-specific lipida transfer protein family. This allergen is resistant to high temperatures, pH changes, gastric and intestinal enzymes. The main route of exposure is through ingestion. In order to examine its resistance to digestion, we have applied a popular 1.0 INFOGEST protocol [1], specialized for the complete food, which in vitro mimics physiologically relevant conditions of oral-gastric-intestinal digestion. The aim of this study was to compare Cor a 8 resistance to gastric digestion, from both, raw and roasted hazelnuts, before and upon pepsin (gastric) digestion. Stability of the Cor a 8 protein was investigated by simulation of oral and gastric digestion phases, performed with ground raw and roasted hazelnut kernels. Hazelnut proteins were extracted from the digestion mixture and analyzed by 1D and 2D SDS-PAGE, while raw and roasted Cor a 8 western blots were probed with specific anti-Cor a 8 antibodies in 1D and 2D immunoblots. The electrophoretic patterns of the raw and roasted extracts were similar. 1D SDS PAGE profiles demonstrated high stability of Cor a 8 against enzymatic treatments. Control samples of Cor a 8 from raw and roasted hazelnut extracts migrated as a single band at around 12 kDa in 1D immunoblot. However, in case of roasted hazelnut, the protein showed a slightly lower capacity to bind specific anti-Cor a 8 antibody, as compared to raw hazelnut extract. In 2D immunoblot, with higher resolution, specific antibody binding was decting a significant and noticeable smear in the basic region indicating a range of different protein variants. This was more pronounced detectable in the case of roasted sample upon digestion, pointing to a mix of variants in this allergen batch. It has been suggested that the allergenicity of the Cor a 8 is almost insensitive to temperature. The allergen is stable even after digestion and roasting processes up to 140˚C. We hypothesize that a lipid-rich food matrix delays extraction of proteins, thereby delaying their gastrointestinal digestion, which may affect allergen sensitizing capacity and clinical symptoms.
PB  - Sociedade Portuguesa de Química
C3  - Book of Abstracts of the XXI EuroFoodChem Congress
T1  - Allergenicity assessment of Cor a 8 from raw and roasted hazelnut upon oral-gastric digestion phase of INFOGEST protocol
EP  - 126
SP  - 126
UR  - https://hdl.handle.net/21.15107/rcub_intor_753
ER  - 

@conference{
author = "Prodić, Ivana and Smiljanić, Katarina and Nagl, Christoph and Hoffmann-Sommergruber, Karin and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Cor a 8 is a relevant allergen that can cause severe allergic reactions. It is a 115 amino acid protein with a molecular mass of 9 kDa and is a member of the non-specific lipida transfer protein family. This allergen is resistant to high temperatures, pH changes, gastric and intestinal enzymes. The main route of exposure is through ingestion. In order to examine its resistance to digestion, we have applied a popular 1.0 INFOGEST protocol [1], specialized for the complete food, which in vitro mimics physiologically relevant conditions of oral-gastric-intestinal digestion. The aim of this study was to compare Cor a 8 resistance to gastric digestion, from both, raw and roasted hazelnuts, before and upon pepsin (gastric) digestion. Stability of the Cor a 8 protein was investigated by simulation of oral and gastric digestion phases, performed with ground raw and roasted hazelnut kernels. Hazelnut proteins were extracted from the digestion mixture and analyzed by 1D and 2D SDS-PAGE, while raw and roasted Cor a 8 western blots were probed with specific anti-Cor a 8 antibodies in 1D and 2D immunoblots. The electrophoretic patterns of the raw and roasted extracts were similar. 1D SDS PAGE profiles demonstrated high stability of Cor a 8 against enzymatic treatments. Control samples of Cor a 8 from raw and roasted hazelnut extracts migrated as a single band at around 12 kDa in 1D immunoblot. However, in case of roasted hazelnut, the protein showed a slightly lower capacity to bind specific anti-Cor a 8 antibody, as compared to raw hazelnut extract. In 2D immunoblot, with higher resolution, specific antibody binding was decting a significant and noticeable smear in the basic region indicating a range of different protein variants. This was more pronounced detectable in the case of roasted sample upon digestion, pointing to a mix of variants in this allergen batch. It has been suggested that the allergenicity of the Cor a 8 is almost insensitive to temperature. The allergen is stable even after digestion and roasting processes up to 140˚C. We hypothesize that a lipid-rich food matrix delays extraction of proteins, thereby delaying their gastrointestinal digestion, which may affect allergen sensitizing capacity and clinical symptoms.",
publisher = "Sociedade Portuguesa de Química",
journal = "Book of Abstracts of the XXI EuroFoodChem Congress",
title = "Allergenicity assessment of Cor a 8 from raw and roasted hazelnut upon oral-gastric digestion phase of INFOGEST protocol",
pages = "126-126",
url = "https://hdl.handle.net/21.15107/rcub_intor_753"
}

Prodić, I., Smiljanić, K., Nagl, C., Hoffmann-Sommergruber, K.,& Ćirković-Veličković, T.. (2021). Allergenicity assessment of Cor a 8 from raw and roasted hazelnut upon oral-gastric digestion phase of INFOGEST protocol. in Book of Abstracts of the XXI EuroFoodChem Congress
Sociedade Portuguesa de Química., 126-126.
https://hdl.handle.net/21.15107/rcub_intor_753

Prodić I, Smiljanić K, Nagl C, Hoffmann-Sommergruber K, Ćirković-Veličković T. Allergenicity assessment of Cor a 8 from raw and roasted hazelnut upon oral-gastric digestion phase of INFOGEST protocol. in Book of Abstracts of the XXI EuroFoodChem Congress. 2021;:126-126.
https://hdl.handle.net/21.15107/rcub_intor_753 .

Prodić, Ivana, Smiljanić, Katarina, Nagl, Christoph, Hoffmann-Sommergruber, Karin, Ćirković-Veličković, Tanja, "Allergenicity assessment of Cor a 8 from raw and roasted hazelnut upon oral-gastric digestion phase of INFOGEST protocol" in Book of Abstracts of the XXI EuroFoodChem Congress (2021):126-126,
https://hdl.handle.net/21.15107/rcub_intor_753 .

TY  - CONF
AU  - Mladenović, Maja
AU  - Romanyuk, Nataliya
AU  - Smiljanić, Katarina
AU  - Jovanović, Vesna B.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/773
AB  - Introduction: Shellfish allergy is one of the most common food allergies with a prevalence of 0.5%-2.5% in the general population. The most common allergen present in shellfish is tropomyosin. Detection of tropomyosin in seashells is a challenge because there are no specific antibodies for seashells’ tropomyosin. Our aim was to verify the presence of tropomyosin in Anadara seashells using an immunoproteomic approach and to investigate the level of cross-reactivity with shrimps.Methods and Results: Proteins from lyophilized seashells Tegillarca granosa (TG) and Anadara broughtonii (AB) were extracted in: RIPA buffer (1% Triton X-100, 1% sodium deoxycholate, 0,1% SDS, 150 mM NaCl, 50 mM Tris-HCl, 1mM EDTA) and Rehydration buffer (7M urea, 2M thiourea, 2% CHAPS and 10mM DTT). Protein concentration of extracts was determined by Bradford assay and SDS-PAGE. The presence of tropomyosin has been supported by commercial tropomyosin standard in 1D SDS-PAGE. With 1D immunoblot, it was possible to confirm the reactivity of seashells’ tropomyosin to rabbit anti-shrimp tropomyosin polyclonal antibodies, confirming its presence. Tropomyosin’s presence was also validated with 1D immunoblot using monoclonal antibodies. 2D electrophoresis showed that most of samples’ proteins are in acidic pI range with prevalence of spots in the range 35-50kDa, and, by comparing spots to 2D immunoblot with polyclonal antibodies, it is possible to confirm tropomyosin’s presence in Anadara seashells.Conclusions: We found that tropomyosin is present in both blood clam species. Both monoclonal and polyclonal antibodies raised against shrimp tropomyosin can detect seashells tropomyosin by immunoblot pointing to a potential antibodies cross-reactivity of allergic subjects to shrimps and seashells.
C3  - FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia
T1  - Detection  and characterization of tropomyosin from Anadara Seashells using immunoproteomic aproach
EP  - 35
SP  - 35
UR  - https://hdl.handle.net/21.15107/rcub_intor_773
ER  - 

@conference{
author = "Mladenović, Maja and Romanyuk, Nataliya and Smiljanić, Katarina and Jovanović, Vesna B. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Introduction: Shellfish allergy is one of the most common food allergies with a prevalence of 0.5%-2.5% in the general population. The most common allergen present in shellfish is tropomyosin. Detection of tropomyosin in seashells is a challenge because there are no specific antibodies for seashells’ tropomyosin. Our aim was to verify the presence of tropomyosin in Anadara seashells using an immunoproteomic approach and to investigate the level of cross-reactivity with shrimps.Methods and Results: Proteins from lyophilized seashells Tegillarca granosa (TG) and Anadara broughtonii (AB) were extracted in: RIPA buffer (1% Triton X-100, 1% sodium deoxycholate, 0,1% SDS, 150 mM NaCl, 50 mM Tris-HCl, 1mM EDTA) and Rehydration buffer (7M urea, 2M thiourea, 2% CHAPS and 10mM DTT). Protein concentration of extracts was determined by Bradford assay and SDS-PAGE. The presence of tropomyosin has been supported by commercial tropomyosin standard in 1D SDS-PAGE. With 1D immunoblot, it was possible to confirm the reactivity of seashells’ tropomyosin to rabbit anti-shrimp tropomyosin polyclonal antibodies, confirming its presence. Tropomyosin’s presence was also validated with 1D immunoblot using monoclonal antibodies. 2D electrophoresis showed that most of samples’ proteins are in acidic pI range with prevalence of spots in the range 35-50kDa, and, by comparing spots to 2D immunoblot with polyclonal antibodies, it is possible to confirm tropomyosin’s presence in Anadara seashells.Conclusions: We found that tropomyosin is present in both blood clam species. Both monoclonal and polyclonal antibodies raised against shrimp tropomyosin can detect seashells tropomyosin by immunoblot pointing to a potential antibodies cross-reactivity of allergic subjects to shrimps and seashells.",
journal = "FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia",
title = "Detection  and characterization of tropomyosin from Anadara Seashells using immunoproteomic aproach",
pages = "35-35",
url = "https://hdl.handle.net/21.15107/rcub_intor_773"
}

Mladenović, M., Romanyuk, N., Smiljanić, K., Jovanović, V. B.,& Ćirković-Veličković, T.. (2021). Detection  and characterization of tropomyosin from Anadara Seashells using immunoproteomic aproach. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia, 35-35.
https://hdl.handle.net/21.15107/rcub_intor_773

Mladenović M, Romanyuk N, Smiljanić K, Jovanović VB, Ćirković-Veličković T. Detection  and characterization of tropomyosin from Anadara Seashells using immunoproteomic aproach. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia. 2021;:35-35.
https://hdl.handle.net/21.15107/rcub_intor_773 .

Mladenović, Maja, Romanyuk, Nataliya, Smiljanić, Katarina, Jovanović, Vesna B., Ćirković-Veličković, Tanja, "Detection  and characterization of tropomyosin from Anadara Seashells using immunoproteomic aproach" in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia (2021):35-35,
https://hdl.handle.net/21.15107/rcub_intor_773 .

TY  - CHAP
AU  - Smiljanić, Katarina
AU  - Mihailović, Jelena
AU  - Prodić, Ivana
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Jovanović, Vesna B.
AU  - Ćirković-Veličković, Tanja
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/764
AB  - Post-translational modifications (PTMs) occur in many forms and shapes, widely influencing protein behavior. High-resolution tandem mass spectrometry (HRMS/MS), coupled with dedicated engines for the identification of unspecified PTMs, is a powerful method for their mapping. A majority of proteomic experiments utilize trypsin for digestion, which cleaves the C-terminal peptide bonds of arginine (Arg) and lysine (Lys) amino acids with high catalytic efficiency and selectivity, unless they are followed with proline. At the same time, Arg and Lys residues are frequently modified during food processing by heat and non-thermal treatments, causing oxidation, carbamylation, and various forms of side chain carbonylation, including the other common PTMs (methylation, acetylation, etc.). Consequently, we explored the possibility to re-assess already generated proteomic data (food protein/allergen tryptic peptides) with respect to the possible modulation of the tryptic intestinal digestion pattern caused by PTMs incorporated at Arg and Lys residues. However, most of the proteomic bottom-up experiments are run with porcine trypsin that has been reductively methylated to increase its stability and minimize autoproteolytic effects. Therefore, in this chapter, the utility of the aforementioned idea was explored, by reviewing the differences in structure, affinity, specificity, and catalytic efficiency of trypsin, primarily from porcine, bovine and human species. Porcine trypsin either from pancreas or in recombinant form showed superior performance compared to human and bovine tryptic counterparts. In addition, set of software tools for identification and analyses of PTMs was reviewed with the aim to isolate those capable of in-depth PTMs profiling and their simultaneous relative quantification, such as PEAKS PTM (PEAKS Studio, Bioinformatics Solution Inc., Ontario Canada). Based on our preliminary experimental results, conclusion is that the proposed idea is plausible, because if potential hindrance effects caused by PTMs are observed with porcine trypsin, then they can be just augmented within human intestinal digestion, with respect to inferior performance of human trypsin.
PB  - New York : Nova Science Publisher
T2  - A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era
T1  - Trypsin as a Proteomic Probe for Assessment of Food Protein Digestibility in Relation to Chemical and Post-translational Modifications
SP  - 158
VL  - 4
UR  - https://hdl.handle.net/21.15107/rcub_intor_764
ER  - 

@inbook{
author = "Smiljanić, Katarina and Mihailović, Jelena and Prodić, Ivana and Đukić, Teodora and Vasović, Tamara and Jovanović, Vesna B. and Ćirković-Veličković, Tanja",
year = "2020",
abstract = "Post-translational modifications (PTMs) occur in many forms and shapes, widely influencing protein behavior. High-resolution tandem mass spectrometry (HRMS/MS), coupled with dedicated engines for the identification of unspecified PTMs, is a powerful method for their mapping. A majority of proteomic experiments utilize trypsin for digestion, which cleaves the C-terminal peptide bonds of arginine (Arg) and lysine (Lys) amino acids with high catalytic efficiency and selectivity, unless they are followed with proline. At the same time, Arg and Lys residues are frequently modified during food processing by heat and non-thermal treatments, causing oxidation, carbamylation, and various forms of side chain carbonylation, including the other common PTMs (methylation, acetylation, etc.). Consequently, we explored the possibility to re-assess already generated proteomic data (food protein/allergen tryptic peptides) with respect to the possible modulation of the tryptic intestinal digestion pattern caused by PTMs incorporated at Arg and Lys residues. However, most of the proteomic bottom-up experiments are run with porcine trypsin that has been reductively methylated to increase its stability and minimize autoproteolytic effects. Therefore, in this chapter, the utility of the aforementioned idea was explored, by reviewing the differences in structure, affinity, specificity, and catalytic efficiency of trypsin, primarily from porcine, bovine and human species. Porcine trypsin either from pancreas or in recombinant form showed superior performance compared to human and bovine tryptic counterparts. In addition, set of software tools for identification and analyses of PTMs was reviewed with the aim to isolate those capable of in-depth PTMs profiling and their simultaneous relative quantification, such as PEAKS PTM (PEAKS Studio, Bioinformatics Solution Inc., Ontario Canada). Based on our preliminary experimental results, conclusion is that the proposed idea is plausible, because if potential hindrance effects caused by PTMs are observed with porcine trypsin, then they can be just augmented within human intestinal digestion, with respect to inferior performance of human trypsin.",
publisher = "New York : Nova Science Publisher",
journal = "A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era",
booktitle = "Trypsin as a Proteomic Probe for Assessment of Food Protein Digestibility in Relation to Chemical and Post-translational Modifications",
pages = "158",
volume = "4",
url = "https://hdl.handle.net/21.15107/rcub_intor_764"
}

Smiljanić, K., Mihailović, J., Prodić, I., Đukić, T., Vasović, T., Jovanović, V. B.,& Ćirković-Veličković, T.. (2020). Trypsin as a Proteomic Probe for Assessment of Food Protein Digestibility in Relation to Chemical and Post-translational Modifications. in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era
New York : Nova Science Publisher., 4, 158.
https://hdl.handle.net/21.15107/rcub_intor_764

Smiljanić K, Mihailović J, Prodić I, Đukić T, Vasović T, Jovanović VB, Ćirković-Veličković T. Trypsin as a Proteomic Probe for Assessment of Food Protein Digestibility in Relation to Chemical and Post-translational Modifications. in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era. 2020;4:158.
https://hdl.handle.net/21.15107/rcub_intor_764 .

Smiljanić, Katarina, Mihailović, Jelena, Prodić, Ivana, Đukić, Teodora, Vasović, Tamara, Jovanović, Vesna B., Ćirković-Veličković, Tanja, "Trypsin as a Proteomic Probe for Assessment of Food Protein Digestibility in Relation to Chemical and Post-translational Modifications" in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era, 4 (2020):158,
https://hdl.handle.net/21.15107/rcub_intor_764 .

TY  - DATA
AU  - Radosavljević, Jelena
AU  - Apostolović, Danijela
AU  - Mihailović, Jelena
AU  - Atanasković-Marković, Marina
AU  - Burazer, Lidija
AU  - van Hage, Marianne
AU  - Ćirković-Veličković, Tanja
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/645
AB  - Figure S1-S3:  Figure S1: Digestion of BLG at pH 1.2, 2.5 and 4.0; Figure S2: Digestion of ALA at pH 1.2, 2.5 and 4.0;  Figure S3: MALDI spectra of peptides used in the study Table S1. IgE levels of patients used in the study determined by ImmunoCAP Methods: 1.1 Detection of ALA and BLG by immunoblot; 1.2 Mass spectrometry analysis; 1.3 Size-exclusion chromatography;  1.4 IgG4-binding properties of peptides obtained by digestion; 1.5 Digestion of purified ALA and BLG at different pH; 1.6 MALDI-TOF MS.
PB  - MDPI
T2  - Foods
T1  - Supplementary information for the article: Radosavljević, J.; Apostolović, D.; Mihailović, J.; Atanasković-Marković, M.;  Burazer, L.; van Hage, M.; Ćirković Veličković, T. Digestomics of Cow’s Milk: Short Digestion-Resistant Peptides of Casein Form  Functional Complexes by Aggregation. Foods 2020, 9 (11), 1576. https://doi.org/10.3390/foods9111576.
IS  - 11
VL  - 9
UR  - https://hdl.handle.net/21.15107/rcub_intor_645
ER  - 

@misc{
author = "Radosavljević, Jelena and Apostolović, Danijela and Mihailović, Jelena and Atanasković-Marković, Marina and Burazer, Lidija and van Hage, Marianne and Ćirković-Veličković, Tanja",
year = "2020",
abstract = "Figure S1-S3:  Figure S1: Digestion of BLG at pH 1.2, 2.5 and 4.0; Figure S2: Digestion of ALA at pH 1.2, 2.5 and 4.0;  Figure S3: MALDI spectra of peptides used in the study Table S1. IgE levels of patients used in the study determined by ImmunoCAP Methods: 1.1 Detection of ALA and BLG by immunoblot; 1.2 Mass spectrometry analysis; 1.3 Size-exclusion chromatography;  1.4 IgG4-binding properties of peptides obtained by digestion; 1.5 Digestion of purified ALA and BLG at different pH; 1.6 MALDI-TOF MS.",
publisher = "MDPI",
journal = "Foods",
title = "Supplementary information for the article: Radosavljević, J.; Apostolović, D.; Mihailović, J.; Atanasković-Marković, M.;  Burazer, L.; van Hage, M.; Ćirković Veličković, T. Digestomics of Cow’s Milk: Short Digestion-Resistant Peptides of Casein Form  Functional Complexes by Aggregation. Foods 2020, 9 (11), 1576. https://doi.org/10.3390/foods9111576.",
number = "11",
volume = "9",
url = "https://hdl.handle.net/21.15107/rcub_intor_645"
}

Radosavljević, J., Apostolović, D., Mihailović, J., Atanasković-Marković, M., Burazer, L., van Hage, M.,& Ćirković-Veličković, T.. (2020). Supplementary information for the article: Radosavljević, J.; Apostolović, D.; Mihailović, J.; Atanasković-Marković, M.;  Burazer, L.; van Hage, M.; Ćirković Veličković, T. Digestomics of Cow’s Milk: Short Digestion-Resistant Peptides of Casein Form  Functional Complexes by Aggregation. Foods 2020, 9 (11), 1576. https://doi.org/10.3390/foods9111576.. in Foods
MDPI., 9(11).
https://hdl.handle.net/21.15107/rcub_intor_645

Radosavljević J, Apostolović D, Mihailović J, Atanasković-Marković M, Burazer L, van Hage M, Ćirković-Veličković T. Supplementary information for the article: Radosavljević, J.; Apostolović, D.; Mihailović, J.; Atanasković-Marković, M.;  Burazer, L.; van Hage, M.; Ćirković Veličković, T. Digestomics of Cow’s Milk: Short Digestion-Resistant Peptides of Casein Form  Functional Complexes by Aggregation. Foods 2020, 9 (11), 1576. https://doi.org/10.3390/foods9111576.. in Foods. 2020;9(11).
https://hdl.handle.net/21.15107/rcub_intor_645 .

Radosavljević, Jelena, Apostolović, Danijela, Mihailović, Jelena, Atanasković-Marković, Marina, Burazer, Lidija, van Hage, Marianne, Ćirković-Veličković, Tanja, "Supplementary information for the article: Radosavljević, J.; Apostolović, D.; Mihailović, J.; Atanasković-Marković, M.;  Burazer, L.; van Hage, M.; Ćirković Veličković, T. Digestomics of Cow’s Milk: Short Digestion-Resistant Peptides of Casein Form  Functional Complexes by Aggregation. Foods 2020, 9 (11), 1576. https://doi.org/10.3390/foods9111576." in Foods, 9, no. 11 (2020),
https://hdl.handle.net/21.15107/rcub_intor_645 .

TY  - JOUR
AU  - Radosavljević, Jelena
AU  - Apostolović, Danijela
AU  - Mihailović, Jelena
AU  - Atanasković-Marković, Marina
AU  - Burazer, Lidija
AU  - van Hage, Marianne
AU  - Ćirković-Veličković, Tanja
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/547
AB  - The aim of this study was to identify short digestion-resistant peptides (SDRPs) released by pepsin digestion of the whole cow's milk and examine their IgE reactivity and allergenicity. Raw milk was subjected to simulated gastric digestion. SDRPs were fractionated from the digests and identified by MS. Milk SDRPs were evaluated for aggregability, propensity to compete for IgE binding with individual milk allergens, and ability to bind IgG4 from allergic and milk-tolerant individuals. The majority of milk SDRPs originated from caseins (97% of peptides) and overlapped with the known IgE epitopes of cow's milk allergens. SDRPs competed with milk proteins for binding to human IgE and readily formed aggregates. The average peptide length was 10.6 +/- 3.5 amino acids. The ability to provoke allergenic in vivo responses was confirmed by skin-prick testing (SPT) in five milk-allergic subjects. This was attributed to the peptide ability to aggregate into non-covalent complexes. SDRPs are able to induce response in SPT, but only in 50% of the sera SDRPs were able to inhibit IgG4 binding to caseins. Hence, SDRPs corresponding to the mainly continuous epitopes of milk proteins induce allergenic in vivo responses in milk-allergic subjects due to aggregation.
PB  - MDPI, Basel
T2  - Foods
T1  - Digestomics of Cow's Milk: Short Digestion-Resistant Peptides of Casein Form Functional Complexes by Aggregation
IS  - 11
VL  - 9
DO  - 10.3390/foods9111576
ER  - 

@article{
author = "Radosavljević, Jelena and Apostolović, Danijela and Mihailović, Jelena and Atanasković-Marković, Marina and Burazer, Lidija and van Hage, Marianne and Ćirković-Veličković, Tanja",
year = "2020",
abstract = "The aim of this study was to identify short digestion-resistant peptides (SDRPs) released by pepsin digestion of the whole cow's milk and examine their IgE reactivity and allergenicity. Raw milk was subjected to simulated gastric digestion. SDRPs were fractionated from the digests and identified by MS. Milk SDRPs were evaluated for aggregability, propensity to compete for IgE binding with individual milk allergens, and ability to bind IgG4 from allergic and milk-tolerant individuals. The majority of milk SDRPs originated from caseins (97% of peptides) and overlapped with the known IgE epitopes of cow's milk allergens. SDRPs competed with milk proteins for binding to human IgE and readily formed aggregates. The average peptide length was 10.6 +/- 3.5 amino acids. The ability to provoke allergenic in vivo responses was confirmed by skin-prick testing (SPT) in five milk-allergic subjects. This was attributed to the peptide ability to aggregate into non-covalent complexes. SDRPs are able to induce response in SPT, but only in 50% of the sera SDRPs were able to inhibit IgG4 binding to caseins. Hence, SDRPs corresponding to the mainly continuous epitopes of milk proteins induce allergenic in vivo responses in milk-allergic subjects due to aggregation.",
publisher = "MDPI, Basel",
journal = "Foods",
title = "Digestomics of Cow's Milk: Short Digestion-Resistant Peptides of Casein Form Functional Complexes by Aggregation",
number = "11",
volume = "9",
doi = "10.3390/foods9111576"
}

Radosavljević, J., Apostolović, D., Mihailović, J., Atanasković-Marković, M., Burazer, L., van Hage, M.,& Ćirković-Veličković, T.. (2020). Digestomics of Cow's Milk: Short Digestion-Resistant Peptides of Casein Form Functional Complexes by Aggregation. in Foods
MDPI, Basel., 9(11).
https://doi.org/10.3390/foods9111576

Radosavljević J, Apostolović D, Mihailović J, Atanasković-Marković M, Burazer L, van Hage M, Ćirković-Veličković T. Digestomics of Cow's Milk: Short Digestion-Resistant Peptides of Casein Form Functional Complexes by Aggregation. in Foods. 2020;9(11).
doi:10.3390/foods9111576 .

Radosavljević, Jelena, Apostolović, Danijela, Mihailović, Jelena, Atanasković-Marković, Marina, Burazer, Lidija, van Hage, Marianne, Ćirković-Veličković, Tanja, "Digestomics of Cow's Milk: Short Digestion-Resistant Peptides of Casein Form Functional Complexes by Aggregation" in Foods, 9, no. 11 (2020),
https://doi.org/10.3390/foods9111576 . .

TY  - CONF
AU  - Peruško, Marija
AU  - Simović, Ana
AU  - Stevanović, Nikola
AU  - Smiljanić, Katarina
AU  - Radomirović, Mirjana Ž.
AU  - Stanić-Vučinić, Dragana
AU  - Ghnimi, Sami
AU  - Ćirković-Veličković, Tanja
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/783
AB  - Objective: Camel milk is highly nutritious food with numerous health benefits proposed. Demandfor camel milk has increased worldwide.Production of camel milk powders facilitate its transport,prolonge shelf-life, and also offer an attractive additive for various food products. In this study wecharacterized proteins of soluble fraction of freeze/spray dried camel milk powders.Material and Methods: Whole camel milk powders were prepared by spray drying treatment at sixdifferent inlet temperatures (190°C - 250°C) or by freeze drying. The soluble protein fractions uponthe treatments were analysed by combination of electrophoretic techniques and circular dichroism.Freeze dried camel milk and spray dried at 250°C were analysed by mass spectrometry.Results: SDS-PAGE revealed non-uniform increase in Mw of major protein bands, while nativeelectrophoresis revealed non-uniform decrease in pI values with increased inlet temperature ofspray drying. That indicated occurence of the Maillard reaction. Far-UV circular dichroism spectrashowed no differences in secondary structures between freeze and spray dried samples. Massspectrometry identified α-lactalbumin, glycosylation-dependant cell adhesion molecule 1(GLYCAM1), immunoglobulin heavy chain, peptidoglycan recognition protein and camel serumalbumin as dominant proteins in soluble fraction of camel milk powders. Carboxymethyl-lisyne(CML), well known marker of Maillard reaction in food analysis, was detected on GLYCAM1 and onimmunoglobulin heavy chain.Conclusions: Our results indicate glycation of camel milk proteins via Maillard reaction upon spraydrying treatment which further may affect techno-functional properties of camel milk powders,their shelf-life and nutritional value.Acknowledgments: This work was supported by the Ministry of Education, Science andTechnological Development of the Republic of Serbia, grant number 172024. The project leading tothis application has received funding from the European Union's Horizon 2020 research andinnovation programme under grant agreement No 810752.
PB  - The Faculty of Sciences, University of Novi Sad, Serbian proteomic association
C3  - The book of abstracts, V SePA symposium: Proteomics in the analysis of food, environmental protection and medical research, 31.5.2019, Novi Sad, Serbia
T1  - Electrophoretic and mass spectrometry-based characterization of soluble fraction of camel milk proteins upon freeze and spray drying treatment
EP  - 7
SP  - 7
UR  - https://hdl.handle.net/21.15107/rcub_intor_783
ER  - 

@conference{
author = "Peruško, Marija and Simović, Ana and Stevanović, Nikola and Smiljanić, Katarina and Radomirović, Mirjana Ž. and Stanić-Vučinić, Dragana and Ghnimi, Sami and Ćirković-Veličković, Tanja",
year = "2019",
abstract = "Objective: Camel milk is highly nutritious food with numerous health benefits proposed. Demandfor camel milk has increased worldwide.Production of camel milk powders facilitate its transport,prolonge shelf-life, and also offer an attractive additive for various food products. In this study wecharacterized proteins of soluble fraction of freeze/spray dried camel milk powders.Material and Methods: Whole camel milk powders were prepared by spray drying treatment at sixdifferent inlet temperatures (190°C - 250°C) or by freeze drying. The soluble protein fractions uponthe treatments were analysed by combination of electrophoretic techniques and circular dichroism.Freeze dried camel milk and spray dried at 250°C were analysed by mass spectrometry.Results: SDS-PAGE revealed non-uniform increase in Mw of major protein bands, while nativeelectrophoresis revealed non-uniform decrease in pI values with increased inlet temperature ofspray drying. That indicated occurence of the Maillard reaction. Far-UV circular dichroism spectrashowed no differences in secondary structures between freeze and spray dried samples. Massspectrometry identified α-lactalbumin, glycosylation-dependant cell adhesion molecule 1(GLYCAM1), immunoglobulin heavy chain, peptidoglycan recognition protein and camel serumalbumin as dominant proteins in soluble fraction of camel milk powders. Carboxymethyl-lisyne(CML), well known marker of Maillard reaction in food analysis, was detected on GLYCAM1 and onimmunoglobulin heavy chain.Conclusions: Our results indicate glycation of camel milk proteins via Maillard reaction upon spraydrying treatment which further may affect techno-functional properties of camel milk powders,their shelf-life and nutritional value.Acknowledgments: This work was supported by the Ministry of Education, Science andTechnological Development of the Republic of Serbia, grant number 172024. The project leading tothis application has received funding from the European Union's Horizon 2020 research andinnovation programme under grant agreement No 810752.",
publisher = "The Faculty of Sciences, University of Novi Sad, Serbian proteomic association",
journal = "The book of abstracts, V SePA symposium: Proteomics in the analysis of food, environmental protection and medical research, 31.5.2019, Novi Sad, Serbia",
title = "Electrophoretic and mass spectrometry-based characterization of soluble fraction of camel milk proteins upon freeze and spray drying treatment",
pages = "7-7",
url = "https://hdl.handle.net/21.15107/rcub_intor_783"
}

Peruško, M., Simović, A., Stevanović, N., Smiljanić, K., Radomirović, M. Ž., Stanić-Vučinić, D., Ghnimi, S.,& Ćirković-Veličković, T.. (2019). Electrophoretic and mass spectrometry-based characterization of soluble fraction of camel milk proteins upon freeze and spray drying treatment. in The book of abstracts, V SePA symposium: Proteomics in the analysis of food, environmental protection and medical research, 31.5.2019, Novi Sad, Serbia
The Faculty of Sciences, University of Novi Sad, Serbian proteomic association., 7-7.
https://hdl.handle.net/21.15107/rcub_intor_783

Peruško M, Simović A, Stevanović N, Smiljanić K, Radomirović MŽ, Stanić-Vučinić D, Ghnimi S, Ćirković-Veličković T. Electrophoretic and mass spectrometry-based characterization of soluble fraction of camel milk proteins upon freeze and spray drying treatment. in The book of abstracts, V SePA symposium: Proteomics in the analysis of food, environmental protection and medical research, 31.5.2019, Novi Sad, Serbia. 2019;:7-7.
https://hdl.handle.net/21.15107/rcub_intor_783 .

Peruško, Marija, Simović, Ana, Stevanović, Nikola, Smiljanić, Katarina, Radomirović, Mirjana Ž., Stanić-Vučinić, Dragana, Ghnimi, Sami, Ćirković-Veličković, Tanja, "Electrophoretic and mass spectrometry-based characterization of soluble fraction of camel milk proteins upon freeze and spray drying treatment" in The book of abstracts, V SePA symposium: Proteomics in the analysis of food, environmental protection and medical research, 31.5.2019, Novi Sad, Serbia (2019):7-7,
https://hdl.handle.net/21.15107/rcub_intor_783 .

TY  - CONF
AU  - Prodić, Ivana
AU  - Smiljanić, Katarina
AU  - Mihailović, Jelena
AU  - Hoffmann-Sommergruber, Karin
AU  - Ćirković-Veličković, Tanja
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/775
AB  - Brief introduction: Stability to gastric digestion represents a very important parameter of food protein allergenicity. Usually digestion experiments are carried out on purified proteins or protein extracts; however, use of solid food is far closer to the in vivo situation, taking into account food protein interactions with other food components, such as polyphenols and lipids.Objective: The aim of this study was to investigate and compare digestion stability and allergenicity of large and small peptides released after pepsin digestion of whole raw and roasted hazelnut kernels under standardized and physiologically relevant in vitro conditions.Methodology: In vitro simulated oral and gastric phase digestion was carried out with ground raw and roasted hazelnut kernels. Digested proteins were extracted from the mixture and analyzed by SDS-PAGE, 2D-PAGE, and compared with Image Master 2D Platinum 7.0. Western blot probed with allergic patients’ sera and specific antibodies for Cor a 8.Main findings: Several important hazelnut seed storage digestion resistant proteins and peptides have been identified and characterized. Most abundant hazelnut allergens were resolved on a 2DE map, for instance acidic and basic chains of Cor a 9, and Cor a11. Digestion-resistant peptides of Cor a 11 and Cor a 9 were able to bind IgE. Lipid transfer protein (Cor a 8) was highly resistant to gastric proteolysis. Conclusion: To conclude, roasted hazelnut is more prone to gastric digestion than raw, and cause milder IgE response in patients. Gastric phase digestion of raw and roasted hazelnut kernels results in partial extraction and digestion of Cor a 11 and Cor a 9 into digestion- resistant peptides with preserved IgE-binding epitopes. These results demonstrate substantial resistance of raw and roasted hazelnut allergens to gastric digestion since they remained mostly intact after 2 h of gastric (pepsin) digestion and retained their allergenicity.
PB  - Univerzitet u Beogradu - Hemijski fakultet
C3  - Abstract Book of the 1st FoodEnTwin Workshop “Food and Environmental -Omics”, Belgrade, June 20-21, 2019
T1  - Digestomics of raw and roasted hazelnut according to Infogest protocol and characterization of gastric-phase products
EP  - 25
SP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_intor_775
ER  - 

@conference{
author = "Prodić, Ivana and Smiljanić, Katarina and Mihailović, Jelena and Hoffmann-Sommergruber, Karin and Ćirković-Veličković, Tanja",
year = "2019",
abstract = "Brief introduction: Stability to gastric digestion represents a very important parameter of food protein allergenicity. Usually digestion experiments are carried out on purified proteins or protein extracts; however, use of solid food is far closer to the in vivo situation, taking into account food protein interactions with other food components, such as polyphenols and lipids.Objective: The aim of this study was to investigate and compare digestion stability and allergenicity of large and small peptides released after pepsin digestion of whole raw and roasted hazelnut kernels under standardized and physiologically relevant in vitro conditions.Methodology: In vitro simulated oral and gastric phase digestion was carried out with ground raw and roasted hazelnut kernels. Digested proteins were extracted from the mixture and analyzed by SDS-PAGE, 2D-PAGE, and compared with Image Master 2D Platinum 7.0. Western blot probed with allergic patients’ sera and specific antibodies for Cor a 8.Main findings: Several important hazelnut seed storage digestion resistant proteins and peptides have been identified and characterized. Most abundant hazelnut allergens were resolved on a 2DE map, for instance acidic and basic chains of Cor a 9, and Cor a11. Digestion-resistant peptides of Cor a 11 and Cor a 9 were able to bind IgE. Lipid transfer protein (Cor a 8) was highly resistant to gastric proteolysis. Conclusion: To conclude, roasted hazelnut is more prone to gastric digestion than raw, and cause milder IgE response in patients. Gastric phase digestion of raw and roasted hazelnut kernels results in partial extraction and digestion of Cor a 11 and Cor a 9 into digestion- resistant peptides with preserved IgE-binding epitopes. These results demonstrate substantial resistance of raw and roasted hazelnut allergens to gastric digestion since they remained mostly intact after 2 h of gastric (pepsin) digestion and retained their allergenicity.",
publisher = "Univerzitet u Beogradu - Hemijski fakultet",
journal = "Abstract Book of the 1st FoodEnTwin Workshop “Food and Environmental -Omics”, Belgrade, June 20-21, 2019",
title = "Digestomics of raw and roasted hazelnut according to Infogest protocol and characterization of gastric-phase products",
pages = "25-25",
url = "https://hdl.handle.net/21.15107/rcub_intor_775"
}

Prodić, I., Smiljanić, K., Mihailović, J., Hoffmann-Sommergruber, K.,& Ćirković-Veličković, T.. (2019). Digestomics of raw and roasted hazelnut according to Infogest protocol and characterization of gastric-phase products. in Abstract Book of the 1st FoodEnTwin Workshop “Food and Environmental -Omics”, Belgrade, June 20-21, 2019
Univerzitet u Beogradu - Hemijski fakultet., 25-25.
https://hdl.handle.net/21.15107/rcub_intor_775

Prodić I, Smiljanić K, Mihailović J, Hoffmann-Sommergruber K, Ćirković-Veličković T. Digestomics of raw and roasted hazelnut according to Infogest protocol and characterization of gastric-phase products. in Abstract Book of the 1st FoodEnTwin Workshop “Food and Environmental -Omics”, Belgrade, June 20-21, 2019. 2019;:25-25.
https://hdl.handle.net/21.15107/rcub_intor_775 .

Prodić, Ivana, Smiljanić, Katarina, Mihailović, Jelena, Hoffmann-Sommergruber, Karin, Ćirković-Veličković, Tanja, "Digestomics of raw and roasted hazelnut according to Infogest protocol and characterization of gastric-phase products" in Abstract Book of the 1st FoodEnTwin Workshop “Food and Environmental -Omics”, Belgrade, June 20-21, 2019 (2019):25-25,
https://hdl.handle.net/21.15107/rcub_intor_775 .

TY  - CONF
AU  - Mihailović, Jelena
AU  - Prodić, Ivana
AU  - Smiljanić, Katarina
AU  - Ćirković-Veličković, Tanja
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/770
AB  - Introduction. Peanut allergy affects a large portion of world population causing reactions rangingfrom mild to severe. Major peanut allergen IgE epitopes are well characterized but little is knownabout their post-translational modifications (PTM) and how they are affected by thermaltreatment. PTM profile may differ between raw and thermally treated peanut, which could affectits allergic potential depending on type, size and position of modifications.Objective. Our aim was to analyse and compare PTM profiles of 4 major peanut allergens - Ara h 1,Ara h 2, Ara h 3 and Ara h 6, as well as their amounts in raw and roasted samples using bottom-upproteomics methods.Methodology. Full peanut protein extracts (both thermally treated and non-treated) were digestedin gel and in solution, and analysed by a Top10 nLC-MS/MS method by LTQ Orbitrap XL (ThermoFisher Scientific Inc., Germany). Within the extracts major allergens - Ara h 1, Ara h 2, Ara h 3 andAra h 6 were identified, label free quantified (LFQ) and searched for PTMs by Peaks X software(Bioinformatics solutions Inc.I, Canada). Epitope sequences were acquired from the ImmuneEpitope Database (IEDB www.iedb.org).Main findings. LFQ results show that there is no significant change in the amountsof any of thestudied allergens between raw and roasted extracts.Out of the 4 allergens Ara h 6 is modified in thehighest portion, with respect to the protein size: 15% and 12% of its positions are modified in rawand roasted sample, respectively. Total of 21 modifications were quantified between the twopreparations, with oxidation (M), methylation (K,R) and dethiomethylation affecting the largestnumber of peptides.Conclusions. Peanut allergen epitopes are indeed carriers of PTMs that differ in pattern andquantity between treated and non-treated extracts. The in silico discovered PTMs could affectprotein digestibility and allergenicity. Further investigation is necessary in order to fully understandthe impact protein modifications could have on their allergenic potential.
PB  - Serbian Proteomic Association - SePA
C3  - Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019
T1  - Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)
SP  - 16/L10
UR  - https://hdl.handle.net/21.15107/rcub_intor_770
ER  - 

@conference{
author = "Mihailović, Jelena and Prodić, Ivana and Smiljanić, Katarina and Ćirković-Veličković, Tanja",
year = "2019",
abstract = "Introduction. Peanut allergy affects a large portion of world population causing reactions rangingfrom mild to severe. Major peanut allergen IgE epitopes are well characterized but little is knownabout their post-translational modifications (PTM) and how they are affected by thermaltreatment. PTM profile may differ between raw and thermally treated peanut, which could affectits allergic potential depending on type, size and position of modifications.Objective. Our aim was to analyse and compare PTM profiles of 4 major peanut allergens - Ara h 1,Ara h 2, Ara h 3 and Ara h 6, as well as their amounts in raw and roasted samples using bottom-upproteomics methods.Methodology. Full peanut protein extracts (both thermally treated and non-treated) were digestedin gel and in solution, and analysed by a Top10 nLC-MS/MS method by LTQ Orbitrap XL (ThermoFisher Scientific Inc., Germany). Within the extracts major allergens - Ara h 1, Ara h 2, Ara h 3 andAra h 6 were identified, label free quantified (LFQ) and searched for PTMs by Peaks X software(Bioinformatics solutions Inc.I, Canada). Epitope sequences were acquired from the ImmuneEpitope Database (IEDB www.iedb.org).Main findings. LFQ results show that there is no significant change in the amountsof any of thestudied allergens between raw and roasted extracts.Out of the 4 allergens Ara h 6 is modified in thehighest portion, with respect to the protein size: 15% and 12% of its positions are modified in rawand roasted sample, respectively. Total of 21 modifications were quantified between the twopreparations, with oxidation (M), methylation (K,R) and dethiomethylation affecting the largestnumber of peptides.Conclusions. Peanut allergen epitopes are indeed carriers of PTMs that differ in pattern andquantity between treated and non-treated extracts. The in silico discovered PTMs could affectprotein digestibility and allergenicity. Further investigation is necessary in order to fully understandthe impact protein modifications could have on their allergenic potential.",
publisher = "Serbian Proteomic Association - SePA",
journal = "Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019",
title = "Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)",
pages = "16/L10",
url = "https://hdl.handle.net/21.15107/rcub_intor_770"
}

Mihailović, J., Prodić, I., Smiljanić, K.,& Ćirković-Veličković, T.. (2019). Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs). in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019
Serbian Proteomic Association - SePA., 16/L10.
https://hdl.handle.net/21.15107/rcub_intor_770

Mihailović J, Prodić I, Smiljanić K, Ćirković-Veličković T. Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs). in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019. 2019;:16/L10.
https://hdl.handle.net/21.15107/rcub_intor_770 .

Mihailović, Jelena, Prodić, Ivana, Smiljanić, Katarina, Ćirković-Veličković, Tanja, "Comparative study of raw and thermally treated peanut major allergen post- translational modifications (PTMs)" in Book of Abstracts - V SePa Simposium: Proteomics in the analysis of food, environmental protection and medical research, Novi Sad 2019 (2019):16/L10,
https://hdl.handle.net/21.15107/rcub_intor_770 .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Apostolović, Danijela
AU  - Cvetković, Anka
AU  - Veljković, Đorđe
AU  - Mutić, Jelena
AU  - van Hage, Marianne
AU  - Burazer, Lidija M.
AU  - Ćirković-Veličković, Tanja
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/763
PB  - Wiley
C3  - Allergy; Congress of the European-Academy-of-Allergy-and-Clinical-Immunology (EAACI)
T1  - In- depth quantitative profiling of post- translational modifications of Timothy grass pollen allergome in relation to environmental pollution and oxidative stress
EP  - 878
IS  - supp. 106
SP  - 878
VL  - 74
UR  - https://hdl.handle.net/21.15107/rcub_intor_763
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana and Apostolović, Danijela and Cvetković, Anka and Veljković, Đorđe and Mutić, Jelena and van Hage, Marianne and Burazer, Lidija M. and Ćirković-Veličković, Tanja",
year = "2019",
publisher = "Wiley",
journal = "Allergy; Congress of the European-Academy-of-Allergy-and-Clinical-Immunology (EAACI)",
title = "In- depth quantitative profiling of post- translational modifications of Timothy grass pollen allergome in relation to environmental pollution and oxidative stress",
pages = "878-878",
number = "supp. 106",
volume = "74",
url = "https://hdl.handle.net/21.15107/rcub_intor_763"
}

Smiljanić, K., Prodić, I., Apostolović, D., Cvetković, A., Veljković, Đ., Mutić, J., van Hage, M., Burazer, L. M.,& Ćirković-Veličković, T.. (2019). In- depth quantitative profiling of post- translational modifications of Timothy grass pollen allergome in relation to environmental pollution and oxidative stress. in Allergy; Congress of the European-Academy-of-Allergy-and-Clinical-Immunology (EAACI)
Wiley., 74(supp. 106), 878-878.
https://hdl.handle.net/21.15107/rcub_intor_763

Smiljanić K, Prodić I, Apostolović D, Cvetković A, Veljković Đ, Mutić J, van Hage M, Burazer LM, Ćirković-Veličković T. In- depth quantitative profiling of post- translational modifications of Timothy grass pollen allergome in relation to environmental pollution and oxidative stress. in Allergy; Congress of the European-Academy-of-Allergy-and-Clinical-Immunology (EAACI). 2019;74(supp. 106):878-878.
https://hdl.handle.net/21.15107/rcub_intor_763 .

Smiljanić, Katarina, Prodić, Ivana, Apostolović, Danijela, Cvetković, Anka, Veljković, Đorđe, Mutić, Jelena, van Hage, Marianne, Burazer, Lidija M., Ćirković-Veličković, Tanja, "In- depth quantitative profiling of post- translational modifications of Timothy grass pollen allergome in relation to environmental pollution and oxidative stress" in Allergy; Congress of the European-Academy-of-Allergy-and-Clinical-Immunology (EAACI), 74, no. supp. 106 (2019):878-878,
https://hdl.handle.net/21.15107/rcub_intor_763 .

TY  - JOUR
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Apostolović, Danijela
AU  - Cvetković, Anka
AU  - Veljović, Đorđe
AU  - Mutić, Jelena
AU  - van Hage, Marianne
AU  - Burazer, Lidija
AU  - Ćirković-Veličković, Tanja
PY  - 2019
UR  - http://intor.torlakinstitut.com/handle/123456789/536
AB  - An association between pollution (e.g., from traffic emissions) and the increased prevalence of respiratory allergies has been observed. Field-realistic exposure studies provide the most relevant assessment of the effects of the intensity and diversity of urban and industrial contamination on pollen structure and allergenicity. The significance of in-depth post-translational modification (PTM) studies of pollen proteomes, when compared with studies on other aspects of pollution and altered pollen allergenicity, has not yet been determined; hence, little progress has been made within this field. We undertook a comprehensive comparative analysis of multiple polluted and environmentally preserved Phleum pratense (Timothy grass) pollen samples using scanning electron microscopy, in-depth PTM profiling, determination of organic and inorganic pollutants, analysis of the release of sub-pollen particles and phenols/proteins, and analysis of proteome expression using high resolution tandem mass spectrometry. In addition, we used quantitative enzyme-linked immunosorbent assays (ELISA) and immunoglobulin E (IgE) immunoblotting. An increased phenolic content and release of sub-pollen particles was found in pollen samples from the polluted area, including a significantly higher content of mercury, cadmium, and manganese, with irregular long spines on pollen grain surface structures. Antioxidative defense-related enzymes were significantly upregulated and seven oxidative PTMs were significantly increased (methionine, histidine, lysine, and proline oxidation; tyrosine glycosylation, lysine 4-hydroxy-2-nonenal adduct, and lysine carbamylation) in pollen exposed to the chemical plant and road traffic pollution sources. Oxidative modifications affected several Timothy pollen allergens; Phl p 6, in particular, exhibited several different oxidative modifications. The expression of Phl p 6, 12, and 13 allergens were downregulated in polluted pollen, and IgE binding to pollen extract was substantially lower in the 18 patients studied, as measured by quantitative ELISA. Quantitative, unrestricted, and detailed PTM searches using an enrichment-free approach pointed to modification of Timothy pollen allergens and suggested that heavy metals are primarily responsible for oxidative stress effects observed in pollen proteins.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Environment International
T1  - In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress
EP  - 658
SP  - 644
VL  - 126
DO  - 10.1016/j.envint.2019.03.001
ER  - 

@article{
author = "Smiljanić, Katarina and Prodić, Ivana and Apostolović, Danijela and Cvetković, Anka and Veljović, Đorđe and Mutić, Jelena and van Hage, Marianne and Burazer, Lidija and Ćirković-Veličković, Tanja",
year = "2019",
abstract = "An association between pollution (e.g., from traffic emissions) and the increased prevalence of respiratory allergies has been observed. Field-realistic exposure studies provide the most relevant assessment of the effects of the intensity and diversity of urban and industrial contamination on pollen structure and allergenicity. The significance of in-depth post-translational modification (PTM) studies of pollen proteomes, when compared with studies on other aspects of pollution and altered pollen allergenicity, has not yet been determined; hence, little progress has been made within this field. We undertook a comprehensive comparative analysis of multiple polluted and environmentally preserved Phleum pratense (Timothy grass) pollen samples using scanning electron microscopy, in-depth PTM profiling, determination of organic and inorganic pollutants, analysis of the release of sub-pollen particles and phenols/proteins, and analysis of proteome expression using high resolution tandem mass spectrometry. In addition, we used quantitative enzyme-linked immunosorbent assays (ELISA) and immunoglobulin E (IgE) immunoblotting. An increased phenolic content and release of sub-pollen particles was found in pollen samples from the polluted area, including a significantly higher content of mercury, cadmium, and manganese, with irregular long spines on pollen grain surface structures. Antioxidative defense-related enzymes were significantly upregulated and seven oxidative PTMs were significantly increased (methionine, histidine, lysine, and proline oxidation; tyrosine glycosylation, lysine 4-hydroxy-2-nonenal adduct, and lysine carbamylation) in pollen exposed to the chemical plant and road traffic pollution sources. Oxidative modifications affected several Timothy pollen allergens; Phl p 6, in particular, exhibited several different oxidative modifications. The expression of Phl p 6, 12, and 13 allergens were downregulated in polluted pollen, and IgE binding to pollen extract was substantially lower in the 18 patients studied, as measured by quantitative ELISA. Quantitative, unrestricted, and detailed PTM searches using an enrichment-free approach pointed to modification of Timothy pollen allergens and suggested that heavy metals are primarily responsible for oxidative stress effects observed in pollen proteins.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Environment International",
title = "In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress",
pages = "658-644",
volume = "126",
doi = "10.1016/j.envint.2019.03.001"
}

Smiljanić, K., Prodić, I., Apostolović, D., Cvetković, A., Veljović, Đ., Mutić, J., van Hage, M., Burazer, L.,& Ćirković-Veličković, T.. (2019). In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress. in Environment International
Pergamon-Elsevier Science Ltd, Oxford., 126, 644-658.
https://doi.org/10.1016/j.envint.2019.03.001

Smiljanić K, Prodić I, Apostolović D, Cvetković A, Veljović Đ, Mutić J, van Hage M, Burazer L, Ćirković-Veličković T. In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress. in Environment International. 2019;126:644-658.
doi:10.1016/j.envint.2019.03.001 .

Smiljanić, Katarina, Prodić, Ivana, Apostolović, Danijela, Cvetković, Anka, Veljović, Đorđe, Mutić, Jelena, van Hage, Marianne, Burazer, Lidija, Ćirković-Veličković, Tanja, "In-depth quantitative profiling of post-translational modifications of Timothy grass pollen allergome in relation to environmental oxidative stress" in Environment International, 126 (2019):644-658,
https://doi.org/10.1016/j.envint.2019.03.001 . .

TY  - CONF
AU  - Prodić, Ivana
AU  - Stanić-Vučinić, Dragana
AU  - Apostolović, Danijela
AU  - Radosavljević, Jelena
AU  - Mihailović, Jelena
AU  - Smiljanić, Katarina
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/789
AB  - Objective: Major peanut allergens, Ara h 2 and Ara h 6, are known to be resistant to pepsindigestion, and they sensitize individual via the gastrointestinal tract. Mikenus et al. published astandardized static digestion method for food, based on physiological conditions emphasizing theimpact of food matrices. Immunoreactive proteins (large fragments) and peptides (short digestionresistant peptides SDRPs; <10 kDa), to which the immune system of the gastrointestinal tract isexposed during digestion of peanut proteins, has not been investigated under pure physiologicalconditions suggested by this protocol.Matherial and methods: Whole grain of grounded raw peanut was incubated with human α-amylase, and pepsin, mimicking the effects of oral and gastric digestion, in total duration of 2h.Bottom up proteomic approach, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, wereused to identify and characterize peanut digesta.Results: After 2h of oral/gastric phase we got, intact proteins, large, digestion resistant peptides(DRP) and SDRPs, as well. Ara h 2 and Ara h 6 remained mostly intact, and short DRPs from Ara h2 and Ara h 6 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h3 showed preserved allergenic capacity, as well. Almost all of identified short DRPs from Ara h 1,Ara h 2 and Ara h 3, with preserved allergenic potential, were constituents of continuous epitopesequences found via Immune Epitope Database (www.iedb.org).Conclusion: Processes of protein extraction from the matrix and their enzymatic digestion occursimultaneously. Oral and gastric phase digestion products of raw peanut are intact proteins, largeand short digestion resistant peptides. Under these conditions Ara h 2 and Ara h 6 are expectedly
PB  - Srpsko Udruženje za Proteomiku; IBISS
C3  - IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
T1  - Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix
UR  - https://hdl.handle.net/21.15107/rcub_intor_789
ER  - 

@conference{
author = "Prodić, Ivana and Stanić-Vučinić, Dragana and Apostolović, Danijela and Radosavljević, Jelena and Mihailović, Jelena and Smiljanić, Katarina and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "Objective: Major peanut allergens, Ara h 2 and Ara h 6, are known to be resistant to pepsindigestion, and they sensitize individual via the gastrointestinal tract. Mikenus et al. published astandardized static digestion method for food, based on physiological conditions emphasizing theimpact of food matrices. Immunoreactive proteins (large fragments) and peptides (short digestionresistant peptides SDRPs; <10 kDa), to which the immune system of the gastrointestinal tract isexposed during digestion of peanut proteins, has not been investigated under pure physiologicalconditions suggested by this protocol.Matherial and methods: Whole grain of grounded raw peanut was incubated with human α-amylase, and pepsin, mimicking the effects of oral and gastric digestion, in total duration of 2h.Bottom up proteomic approach, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, wereused to identify and characterize peanut digesta.Results: After 2h of oral/gastric phase we got, intact proteins, large, digestion resistant peptides(DRP) and SDRPs, as well. Ara h 2 and Ara h 6 remained mostly intact, and short DRPs from Ara h2 and Ara h 6 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h3 showed preserved allergenic capacity, as well. Almost all of identified short DRPs from Ara h 1,Ara h 2 and Ara h 3, with preserved allergenic potential, were constituents of continuous epitopesequences found via Immune Epitope Database (www.iedb.org).Conclusion: Processes of protein extraction from the matrix and their enzymatic digestion occursimultaneously. Oral and gastric phase digestion products of raw peanut are intact proteins, largeand short digestion resistant peptides. Under these conditions Ara h 2 and Ara h 6 are expectedly",
publisher = "Srpsko Udruženje za Proteomiku; IBISS",
journal = "IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija",
title = "Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix",
url = "https://hdl.handle.net/21.15107/rcub_intor_789"
}

Prodić, I., Stanić-Vučinić, D., Apostolović, D., Radosavljević, J., Mihailović, J., Smiljanić, K.,& Ćirković-Veličković, T.. (2018). Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
Srpsko Udruženje za Proteomiku; IBISS..
https://hdl.handle.net/21.15107/rcub_intor_789

Prodić I, Stanić-Vučinić D, Apostolović D, Radosavljević J, Mihailović J, Smiljanić K, Ćirković-Veličković T. Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija. 2018;.
https://hdl.handle.net/21.15107/rcub_intor_789 .

Prodić, Ivana, Stanić-Vučinić, Dragana, Apostolović, Danijela, Radosavljević, Jelena, Mihailović, Jelena, Smiljanić, Katarina, Ćirković-Veličković, Tanja, "Gastric digestome of whole peanut grains from the aspect of immunoproteomics: Characterization of digested allergens in the real food matrix" in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija (2018),
https://hdl.handle.net/21.15107/rcub_intor_789 .

TY  - CONF
AU  - Prodić, Ivana
AU  - Khulal, Urmila
AU  - Mutić, Jelena
AU  - Mihailović, Jelena
AU  - Smiljanić, Katarina
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/774
AB  - Objective: Haliotis discus (Japanese abalone), mollusks among various shellfish, is a highlynutritive food resource in the world, but also among the eight allergic food groups accounting forapproximately 90% of all immunoglobulin E food allergies worldwide. The general objective of ourresearch is to comprehensively investigate stability and structures of pepsin-resistant allergens, oftheir larger fragments, and of short digestion resistant peptides (SDRPs) released by pepsindigestion of whole raw and extract of shellfish, under standardized and physiologically relevantgastric conditions.Materials and Methods: Extract of raw whole shellfish (eRSS) and whole raw shellfish (wRSS),were pepsin digested according to standardized static digestion protocol. Controls were treated in asame manner without adding pepsin. Supernatant of samples and its counterpart controls wereprecipitated with TCA/acetone. Obtained proteins were assessed by 2D SDS PAGE and 1D SDS-PAGE, under reducing and non-reducing conditions. 1D SDS-PAGE of RSS were analyzed byncLC-MS/MS (Orbitrap LTQ) shot-gun proteomics. Relative quantification was performed by LFQalgorithm within Peaks 8.5 software package Bioinformatics Solutions Inc. (BSI), Waterloo,Canada.Results and Conclusion: 1D SDS-PAGE analysis of eRSS and wRSS, and its controls showed arange of proteins in varied concentrations between 10-250 kDa. In extracted and whole rawshellfish, approximately 22 prominent protein bands were observed including the distinct bandscorresponding with the molecular weights of recognized shellfish allergen, tropomyosin (37-39kDa). Fewer high molecular weight proteins were observed followed by protein smearing,specifically around the low molecular weight protein bands. The smearing could possibly be due tothe breakdown products and the glycation. There were slight differences between the proteinprofiles under reducing and non-reducing conditions as well. Nevertheless, there was the retentionof a band in the 37kDa molecular weight marker in all 4 samples, likely consistent with heat stabletropomyosin (TM). Mass spectrometry showed allergens that are characterized (Hal d 1 and Hal di1), with 90% of sequence homology with main tropomyosin allergens from seafood.Scientific impact and relevance: The results will highlight effects of food matrix on shellfishallergens digestibility proving its relevancy in molecular allergology. Moreover, an insight will beobtained on the differences in digestibility of allergenic versus non-allergenic tropomyosins in thereal food matrix.
PB  - Srpsko Udruženje za Proteomiku, SePA; IBISS
C3  - IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
T1  - Digestomics of Japanese abalone in real food matrix
EP  - 10
SP  - 10
UR  - https://hdl.handle.net/21.15107/rcub_intor_774
ER  - 

@conference{
author = "Prodić, Ivana and Khulal, Urmila and Mutić, Jelena and Mihailović, Jelena and Smiljanić, Katarina and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "Objective: Haliotis discus (Japanese abalone), mollusks among various shellfish, is a highlynutritive food resource in the world, but also among the eight allergic food groups accounting forapproximately 90% of all immunoglobulin E food allergies worldwide. The general objective of ourresearch is to comprehensively investigate stability and structures of pepsin-resistant allergens, oftheir larger fragments, and of short digestion resistant peptides (SDRPs) released by pepsindigestion of whole raw and extract of shellfish, under standardized and physiologically relevantgastric conditions.Materials and Methods: Extract of raw whole shellfish (eRSS) and whole raw shellfish (wRSS),were pepsin digested according to standardized static digestion protocol. Controls were treated in asame manner without adding pepsin. Supernatant of samples and its counterpart controls wereprecipitated with TCA/acetone. Obtained proteins were assessed by 2D SDS PAGE and 1D SDS-PAGE, under reducing and non-reducing conditions. 1D SDS-PAGE of RSS were analyzed byncLC-MS/MS (Orbitrap LTQ) shot-gun proteomics. Relative quantification was performed by LFQalgorithm within Peaks 8.5 software package Bioinformatics Solutions Inc. (BSI), Waterloo,Canada.Results and Conclusion: 1D SDS-PAGE analysis of eRSS and wRSS, and its controls showed arange of proteins in varied concentrations between 10-250 kDa. In extracted and whole rawshellfish, approximately 22 prominent protein bands were observed including the distinct bandscorresponding with the molecular weights of recognized shellfish allergen, tropomyosin (37-39kDa). Fewer high molecular weight proteins were observed followed by protein smearing,specifically around the low molecular weight protein bands. The smearing could possibly be due tothe breakdown products and the glycation. There were slight differences between the proteinprofiles under reducing and non-reducing conditions as well. Nevertheless, there was the retentionof a band in the 37kDa molecular weight marker in all 4 samples, likely consistent with heat stabletropomyosin (TM). Mass spectrometry showed allergens that are characterized (Hal d 1 and Hal di1), with 90% of sequence homology with main tropomyosin allergens from seafood.Scientific impact and relevance: The results will highlight effects of food matrix on shellfishallergens digestibility proving its relevancy in molecular allergology. Moreover, an insight will beobtained on the differences in digestibility of allergenic versus non-allergenic tropomyosins in thereal food matrix.",
publisher = "Srpsko Udruženje za Proteomiku, SePA; IBISS",
journal = "IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija",
title = "Digestomics of Japanese abalone in real food matrix",
pages = "10-10",
url = "https://hdl.handle.net/21.15107/rcub_intor_774"
}

Prodić, I., Khulal, U., Mutić, J., Mihailović, J., Smiljanić, K.,& Ćirković-Veličković, T.. (2018). Digestomics of Japanese abalone in real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
Srpsko Udruženje za Proteomiku, SePA; IBISS., 10-10.
https://hdl.handle.net/21.15107/rcub_intor_774

Prodić I, Khulal U, Mutić J, Mihailović J, Smiljanić K, Ćirković-Veličković T. Digestomics of Japanese abalone in real food matrix. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija. 2018;:10-10.
https://hdl.handle.net/21.15107/rcub_intor_774 .

Prodić, Ivana, Khulal, Urmila, Mutić, Jelena, Mihailović, Jelena, Smiljanić, Katarina, Ćirković-Veličković, Tanja, "Digestomics of Japanese abalone in real food matrix" in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija (2018):10-10,
https://hdl.handle.net/21.15107/rcub_intor_774 .

TY  - CONF
AU  - Prodić, Ivana
AU  - Dubiela, Pawel
AU  - Mihailović, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Smiljanić, Katarina
AU  - Hoffmann-Sommergruber, Karin
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/776
AB  - Background: Sensitization to non-specific lipid transfer protein (ns-LTPs) in plant foods is regarded as a risk factor for generalized allergic reactions. Stability to gastric digestion represents very important parameter of food proteins allergenicity. Usually studies of digestion were carried out on purified proteins, but has never been examined the influence of different food matrices on different allergens. Allergens from the nsLTP family are known to share a characteristic structure which is highly resistant to proteolysis, and therefore, IgE cross-reactivity of nsLTPs needs to be investigated in the environment such as complex food matrix.Objective: The aim of this research project is to reveal how proteins are digested (by Minekus protocol) within the natural food matrix and possible consequences on their allergenicity, with the special focus on ns-LTP.Methods: Pure nsLTPs from walnut were labelled with Alexa 633 and added to whole grain of grounded raw walnuts, incubated with human α-amylase, and pepsin, therefore mimicking the effects of oral and gastric digestion, in total duration of 2h. Proteins extracted from the mixture were analyzed by one-dimensional (1D) and two-dimensional SDS-PAGe, and respective 1D and 2D immunoblots.Results: Most proteins from pepsin digested walnut sample were more resistant to digestion according to 1D SDS PAGE. Pepsin digested raw walnut sample with nsLTP were assessed by 2D PAGE to compare profiles of the digested and control sample (no pepsin added). 2D SDSPAGE of digested and control walnut samples showed almost identical profiles, especially in the context of fluorescently labelled nsLTP allergens. These results demonstrate substantial resistance of nsLTP allergens to gastric digestion since they remained mostly intact after 2 h of gastric (pepsin) digestion.Conclusion: Further research is needed to be able to grade stability/resistance of selected food allergens to gastric digestion as a consequence of food matrix modulating effects. We propose that certain combinations of foods and allergens could provide additional protection or on the contrary ease the digestion, by comparing trends between control and digested samples and between different digested combinations as well.
PB  - IMPARAS Cost Action FA1402
C3  - Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018
T1  - Digestomics of walnut and its nsLTPs allergens reveals their ultimate resistance to gastric digestion
EP  - 59
SP  - 59
UR  - https://hdl.handle.net/21.15107/rcub_intor_776
ER  - 

@conference{
author = "Prodić, Ivana and Dubiela, Pawel and Mihailović, Jelena and Stanić-Vučinić, Dragana and Smiljanić, Katarina and Hoffmann-Sommergruber, Karin and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "Background: Sensitization to non-specific lipid transfer protein (ns-LTPs) in plant foods is regarded as a risk factor for generalized allergic reactions. Stability to gastric digestion represents very important parameter of food proteins allergenicity. Usually studies of digestion were carried out on purified proteins, but has never been examined the influence of different food matrices on different allergens. Allergens from the nsLTP family are known to share a characteristic structure which is highly resistant to proteolysis, and therefore, IgE cross-reactivity of nsLTPs needs to be investigated in the environment such as complex food matrix.Objective: The aim of this research project is to reveal how proteins are digested (by Minekus protocol) within the natural food matrix and possible consequences on their allergenicity, with the special focus on ns-LTP.Methods: Pure nsLTPs from walnut were labelled with Alexa 633 and added to whole grain of grounded raw walnuts, incubated with human α-amylase, and pepsin, therefore mimicking the effects of oral and gastric digestion, in total duration of 2h. Proteins extracted from the mixture were analyzed by one-dimensional (1D) and two-dimensional SDS-PAGe, and respective 1D and 2D immunoblots.Results: Most proteins from pepsin digested walnut sample were more resistant to digestion according to 1D SDS PAGE. Pepsin digested raw walnut sample with nsLTP were assessed by 2D PAGE to compare profiles of the digested and control sample (no pepsin added). 2D SDSPAGE of digested and control walnut samples showed almost identical profiles, especially in the context of fluorescently labelled nsLTP allergens. These results demonstrate substantial resistance of nsLTP allergens to gastric digestion since they remained mostly intact after 2 h of gastric (pepsin) digestion.Conclusion: Further research is needed to be able to grade stability/resistance of selected food allergens to gastric digestion as a consequence of food matrix modulating effects. We propose that certain combinations of foods and allergens could provide additional protection or on the contrary ease the digestion, by comparing trends between control and digested samples and between different digested combinations as well.",
publisher = "IMPARAS Cost Action FA1402",
journal = "Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018",
title = "Digestomics of walnut and its nsLTPs allergens reveals their ultimate resistance to gastric digestion",
pages = "59-59",
url = "https://hdl.handle.net/21.15107/rcub_intor_776"
}

Prodić, I., Dubiela, P., Mihailović, J., Stanić-Vučinić, D., Smiljanić, K., Hoffmann-Sommergruber, K.,& Ćirković-Veličković, T.. (2018). Digestomics of walnut and its nsLTPs allergens reveals their ultimate resistance to gastric digestion. in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018
IMPARAS Cost Action FA1402., 59-59.
https://hdl.handle.net/21.15107/rcub_intor_776

Prodić I, Dubiela P, Mihailović J, Stanić-Vučinić D, Smiljanić K, Hoffmann-Sommergruber K, Ćirković-Veličković T. Digestomics of walnut and its nsLTPs allergens reveals their ultimate resistance to gastric digestion. in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018. 2018;:59-59.
https://hdl.handle.net/21.15107/rcub_intor_776 .

Prodić, Ivana, Dubiela, Pawel, Mihailović, Jelena, Stanić-Vučinić, Dragana, Smiljanić, Katarina, Hoffmann-Sommergruber, Karin, Ćirković-Veličković, Tanja, "Digestomics of walnut and its nsLTPs allergens reveals their ultimate resistance to gastric digestion" in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018 (2018):59-59,
https://hdl.handle.net/21.15107/rcub_intor_776 .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Aleksić, Ivana
AU  - Veljović, Đorđe
AU  - Ćirković-Veličković, Tanja
AU  - Mutić, Jelena
AU  - Burazer, Lidija M.
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/772
AB  - Objective: to create method for unrestrictive deep, relative quantification of post translationalmodifications (PTMs) within different proteomes. Pollution field studies of bio indicators such aspollen are valuable because of realistic situation of target contamination, however they carry thegreat extent of uncertainty in attributing and delineating the polluting effect from multiple sources.Holistic research platform focusing on comprehensively characterized and quantified PTMs ofcomparable bio-indicator proteomes could help and overcome these obstacles of field pollutionstudies.Material and Methods: Scanning electron and light microscopy assessed surface and sub pollenparticle (SPP) releasing features of timothy grass (TG) pollen. Inductively coupled atomic emissionspectrometry revealed metal elemental content in pollen while in solution trypsin digested pollenproteomes analysed with high resolution Orbitrap mass tandem spectrometry and PEAKS Suite 8.5brought quantitative information on protein expression level and its PTM profiling.Results: TG polluted pollen samples (P2) collected along regional road and chemical plant,exposed to air contaminants from road traffics and chemical plants showed 4.5 times higher SPPreleasing capacity, with notable surface changes, as well as significantly higher contents of Mn, Hgand Cd. Antioxidative enzymes (oxidoreductases, superoxide dismutases and peroxidases),including actin, were upregulated several times in polluted sample compared to ecologicallypreserved pollen (P1). While the level of spontaneous and physiological PTMs includingmethylation, acetylation, deamidation and formylation, was similar without significant changes inP1 and P2 pollens, oxidative PTMs including oxidation of Met, Lys, His, Pro and HNE and hexoseadducts showed several times higher and significant increase in abundancy of P2 compared to P1.PTMs connected to road traffic such as tyrosine nitration were very rare and low abundant.Conclusion: Results suggest prominent role of chemical pollution compared to effect of road trafficpollution, with primary consequences from oxidative properties of mercury (Hg) and cadmium(Cd).
PB  - Srpsko Udruženje za Proteomiku, SePA; IBISS
C3  - IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
T1  - Deep and quantitative profiling of PTMs in ecologically preserved and polluted pollen proteomes of timothy grass reveals predominant source of contamination
EP  - 13
SP  - 13
UR  - https://hdl.handle.net/21.15107/rcub_intor_772
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana and Aleksić, Ivana and Veljović, Đorđe and Ćirković-Veličković, Tanja and Mutić, Jelena and Burazer, Lidija M.",
year = "2018",
abstract = "Objective: to create method for unrestrictive deep, relative quantification of post translationalmodifications (PTMs) within different proteomes. Pollution field studies of bio indicators such aspollen are valuable because of realistic situation of target contamination, however they carry thegreat extent of uncertainty in attributing and delineating the polluting effect from multiple sources.Holistic research platform focusing on comprehensively characterized and quantified PTMs ofcomparable bio-indicator proteomes could help and overcome these obstacles of field pollutionstudies.Material and Methods: Scanning electron and light microscopy assessed surface and sub pollenparticle (SPP) releasing features of timothy grass (TG) pollen. Inductively coupled atomic emissionspectrometry revealed metal elemental content in pollen while in solution trypsin digested pollenproteomes analysed with high resolution Orbitrap mass tandem spectrometry and PEAKS Suite 8.5brought quantitative information on protein expression level and its PTM profiling.Results: TG polluted pollen samples (P2) collected along regional road and chemical plant,exposed to air contaminants from road traffics and chemical plants showed 4.5 times higher SPPreleasing capacity, with notable surface changes, as well as significantly higher contents of Mn, Hgand Cd. Antioxidative enzymes (oxidoreductases, superoxide dismutases and peroxidases),including actin, were upregulated several times in polluted sample compared to ecologicallypreserved pollen (P1). While the level of spontaneous and physiological PTMs includingmethylation, acetylation, deamidation and formylation, was similar without significant changes inP1 and P2 pollens, oxidative PTMs including oxidation of Met, Lys, His, Pro and HNE and hexoseadducts showed several times higher and significant increase in abundancy of P2 compared to P1.PTMs connected to road traffic such as tyrosine nitration were very rare and low abundant.Conclusion: Results suggest prominent role of chemical pollution compared to effect of road trafficpollution, with primary consequences from oxidative properties of mercury (Hg) and cadmium(Cd).",
publisher = "Srpsko Udruženje za Proteomiku, SePA; IBISS",
journal = "IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija",
title = "Deep and quantitative profiling of PTMs in ecologically preserved and polluted pollen proteomes of timothy grass reveals predominant source of contamination",
pages = "13-13",
url = "https://hdl.handle.net/21.15107/rcub_intor_772"
}

Smiljanić, K., Prodić, I., Aleksić, I., Veljović, Đ., Ćirković-Veličković, T., Mutić, J.,& Burazer, L. M.. (2018). Deep and quantitative profiling of PTMs in ecologically preserved and polluted pollen proteomes of timothy grass reveals predominant source of contamination. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija
Srpsko Udruženje za Proteomiku, SePA; IBISS., 13-13.
https://hdl.handle.net/21.15107/rcub_intor_772

Smiljanić K, Prodić I, Aleksić I, Veljović Đ, Ćirković-Veličković T, Mutić J, Burazer LM. Deep and quantitative profiling of PTMs in ecologically preserved and polluted pollen proteomes of timothy grass reveals predominant source of contamination. in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija. 2018;:13-13.
https://hdl.handle.net/21.15107/rcub_intor_772 .

Smiljanić, Katarina, Prodić, Ivana, Aleksić, Ivana, Veljović, Đorđe, Ćirković-Veličković, Tanja, Mutić, Jelena, Burazer, Lidija M., "Deep and quantitative profiling of PTMs in ecologically preserved and polluted pollen proteomes of timothy grass reveals predominant source of contamination" in IV Simpozijum srpskog udruženja za proteomiku – SePA, Interaktomika i glikoproteomika: novi pristup u analizi proteina na velikoj skali, 25. maj 2018, Beograd, Srbija (2018):13-13,
https://hdl.handle.net/21.15107/rcub_intor_772 .

TY  - CONF
AU  - Liu, Shu-hua
AU  - Mihailović, Jelena
AU  - Smiljanić, Katarina
AU  - Epstein, Michelle M.
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/768
AB  - BackgroundPeanut allergy is the most common type of food allergy causing severe reactions or even fatal anaphylaxis in sensitised individuals. The major peanut allergens are Ara h 1, Ara h 2, Ara h 3, and Ara h 6 which cause the most severe responses. Their molecular properties have been characterised but possible post-translational modifications (PTMs) that might explain their severe allergenicity are not well understood. The goal of this study was to utilize a combination of nanoLC-Mass Spectrometry (MS)/MS methods and PEAKS Studio 8.0 (Bioinformatics Solutions Inc., Ontario, Canada) program to evaluate PTMs in the major peanut allergens.MethodAcquired MS data of purified peanut allergens, Ara h 1, Ara h 2, Ara h 3, and Ara h 6 were analysed and identified via hybridized databases obtained from UniProt (www.uniprot.org).More than 1200 reviewed (Swiss-Prot) and unreviewed (TrEMBL) entries from peanut were combined with common MS contaminants, the Repository of Adventitious Proteins (cRAP), to create a hybridized database. We then focused on Ara h 2 (Conglutin-7) and Ara h 6 (Conglutin) because of their propensity to cause severe anaphylactic reactions. Epitopes found in the Immune Epitope Database (www.iedb.org) were analysed for possible PTMs by matching PEAKS PTM results with mapped positions of epitope sequences.ResultsWe identified 37 proteins from the purified peanut allergens. There were 33 peanut proteins and 4 contaminants originating from human keratin and pig trypsin. Ara h 2 had 242 epitopes, 29 potential PTMs and 4 mutations. Eight of the epitopes had up to 8 possible PTMs. Several relevant PTMs were discovered, including tryptophan oxidation to oxolactone in position 25, sulfonation of N-terminus of cysteine in position 116 and oxidation of methionine in position 50 and 125. Notably, all had either a “NNQRCMCEALQ” or “QQIMENQSD” motif, which are linked to Th2 cytokines and T cell proliferation. We observed 8 epitopes, 9 likely PTMs and no mutations for Ara h 6 and half of the epitopes had possible PTMs and a maximum of 4 PTMs was found on one epitope.ConclusionThe analysis of relevant peanut allergens by nanoLC-MS/MS methods and PEAKS Studio 8.0 program revealed several PTMs, which might have important ramifications due to their influence on allergenicity and digestibility resulting from modification properties by trypsin and other food protein enzymes. These data suggest that PTMs on certain peanut epitopes could be involved in the pathogenesis of severe food allergy to peanuts.
PB  - IMPARAS Cost Action FA1402
C3  - Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018
T1  - Characterisation of peanut allergens and possible post-translational modifications (PTMs)
EP  - 57
SP  - 57
UR  - https://hdl.handle.net/21.15107/rcub_intor_768
ER  - 

@conference{
author = "Liu, Shu-hua and Mihailović, Jelena and Smiljanić, Katarina and Epstein, Michelle M. and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "BackgroundPeanut allergy is the most common type of food allergy causing severe reactions or even fatal anaphylaxis in sensitised individuals. The major peanut allergens are Ara h 1, Ara h 2, Ara h 3, and Ara h 6 which cause the most severe responses. Their molecular properties have been characterised but possible post-translational modifications (PTMs) that might explain their severe allergenicity are not well understood. The goal of this study was to utilize a combination of nanoLC-Mass Spectrometry (MS)/MS methods and PEAKS Studio 8.0 (Bioinformatics Solutions Inc., Ontario, Canada) program to evaluate PTMs in the major peanut allergens.MethodAcquired MS data of purified peanut allergens, Ara h 1, Ara h 2, Ara h 3, and Ara h 6 were analysed and identified via hybridized databases obtained from UniProt (www.uniprot.org).More than 1200 reviewed (Swiss-Prot) and unreviewed (TrEMBL) entries from peanut were combined with common MS contaminants, the Repository of Adventitious Proteins (cRAP), to create a hybridized database. We then focused on Ara h 2 (Conglutin-7) and Ara h 6 (Conglutin) because of their propensity to cause severe anaphylactic reactions. Epitopes found in the Immune Epitope Database (www.iedb.org) were analysed for possible PTMs by matching PEAKS PTM results with mapped positions of epitope sequences.ResultsWe identified 37 proteins from the purified peanut allergens. There were 33 peanut proteins and 4 contaminants originating from human keratin and pig trypsin. Ara h 2 had 242 epitopes, 29 potential PTMs and 4 mutations. Eight of the epitopes had up to 8 possible PTMs. Several relevant PTMs were discovered, including tryptophan oxidation to oxolactone in position 25, sulfonation of N-terminus of cysteine in position 116 and oxidation of methionine in position 50 and 125. Notably, all had either a “NNQRCMCEALQ” or “QQIMENQSD” motif, which are linked to Th2 cytokines and T cell proliferation. We observed 8 epitopes, 9 likely PTMs and no mutations for Ara h 6 and half of the epitopes had possible PTMs and a maximum of 4 PTMs was found on one epitope.ConclusionThe analysis of relevant peanut allergens by nanoLC-MS/MS methods and PEAKS Studio 8.0 program revealed several PTMs, which might have important ramifications due to their influence on allergenicity and digestibility resulting from modification properties by trypsin and other food protein enzymes. These data suggest that PTMs on certain peanut epitopes could be involved in the pathogenesis of severe food allergy to peanuts.",
publisher = "IMPARAS Cost Action FA1402",
journal = "Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018",
title = "Characterisation of peanut allergens and possible post-translational modifications (PTMs)",
pages = "57-57",
url = "https://hdl.handle.net/21.15107/rcub_intor_768"
}

Liu, S., Mihailović, J., Smiljanić, K., Epstein, M. M.,& Ćirković-Veličković, T.. (2018). Characterisation of peanut allergens and possible post-translational modifications (PTMs). in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018
IMPARAS Cost Action FA1402., 57-57.
https://hdl.handle.net/21.15107/rcub_intor_768

Liu S, Mihailović J, Smiljanić K, Epstein MM, Ćirković-Veličković T. Characterisation of peanut allergens and possible post-translational modifications (PTMs). in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018. 2018;:57-57.
https://hdl.handle.net/21.15107/rcub_intor_768 .

Liu, Shu-hua, Mihailović, Jelena, Smiljanić, Katarina, Epstein, Michelle M., Ćirković-Veličković, Tanja, "Characterisation of peanut allergens and possible post-translational modifications (PTMs)" in Proceedings of the 4th International ImpARAS Conference, Portici (Naples), Italy, June 19-21, 2018 (2018):57-57,
https://hdl.handle.net/21.15107/rcub_intor_768 .

TY  - JOUR
AU  - Prodić, Ivana
AU  - Stanić-Vučinić, Dragana
AU  - Apostolović, Danijela
AU  - Mihailović, Jelena
AU  - Radibratović, M.
AU  - Radosavljević, Jelena
AU  - Burazer, Lidija
AU  - Milcić, M.
AU  - Smiljanić, Katarina
AU  - van Hage, Marianne
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/608
AB  - Background: Most food allergens sensitizing via the gastrointestinal tract are stable proteins that are resistant to pepsin digestion, in particular major peanut allergens, Ara h 2 and Ara h 6. Survival of their large fragments is essential for sensitizing capacity. However, the immunoreactive proteins/peptides to which the immune system of the gastrointestinal tract is exposed during digestion of peanut proteins are unknown. Particularly, the IgE reactivity of short digestion-resistant peptides (SDRPs;  lt 10 kDa) released by gastric digestion under standardized and physiologically relevant in vitro conditions has not been investigated. Objective: The aim of this study was to investigate and identify digestion products of major peanut allergens and in particular to examine IgE reactivity of SDRPs released by pepsin digestion of whole peanut grains. Methods: Two-dimensional gel-based proteomics and shotgun peptidomics, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, including far ultraviolet-circular dichroism spectroscopy were used to identify and characterize peanut digesta. Results: Ara h 2 and Ara h 6 remained mostly intact, and SDRPs from Ara h 2 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h 3 exhibited sequential digestion into a series of digestion-resistant peptides with preserved allergenic capacity. A high number of identified SDRPs from Ara h 1, Ara h 2 and Ara h 3 were part of short continuous epitope sequences and possessed substantial allergenic potential. Conclusion and Clinical Relevance: Peanut grain digestion by oral and gastric phase enzymes generates mixture of products, where the major peanut allergens remain intact and their digested peptides have preserved allergenic capacity highlighting their important roles in allergic reactions to peanut.
PB  - Blackwell Publishing Ltd
T2  - Clinical and Experimental Allergy
T1  - Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides
EP  - 740
IS  - 6
SP  - 731
VL  - 48
DO  - 10.1111/cea.13113
ER  - 

@article{
author = "Prodić, Ivana and Stanić-Vučinić, Dragana and Apostolović, Danijela and Mihailović, Jelena and Radibratović, M. and Radosavljević, Jelena and Burazer, Lidija and Milcić, M. and Smiljanić, Katarina and van Hage, Marianne and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "Background: Most food allergens sensitizing via the gastrointestinal tract are stable proteins that are resistant to pepsin digestion, in particular major peanut allergens, Ara h 2 and Ara h 6. Survival of their large fragments is essential for sensitizing capacity. However, the immunoreactive proteins/peptides to which the immune system of the gastrointestinal tract is exposed during digestion of peanut proteins are unknown. Particularly, the IgE reactivity of short digestion-resistant peptides (SDRPs;  lt 10 kDa) released by gastric digestion under standardized and physiologically relevant in vitro conditions has not been investigated. Objective: The aim of this study was to investigate and identify digestion products of major peanut allergens and in particular to examine IgE reactivity of SDRPs released by pepsin digestion of whole peanut grains. Methods: Two-dimensional gel-based proteomics and shotgun peptidomics, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, including far ultraviolet-circular dichroism spectroscopy were used to identify and characterize peanut digesta. Results: Ara h 2 and Ara h 6 remained mostly intact, and SDRPs from Ara h 2 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h 3 exhibited sequential digestion into a series of digestion-resistant peptides with preserved allergenic capacity. A high number of identified SDRPs from Ara h 1, Ara h 2 and Ara h 3 were part of short continuous epitope sequences and possessed substantial allergenic potential. Conclusion and Clinical Relevance: Peanut grain digestion by oral and gastric phase enzymes generates mixture of products, where the major peanut allergens remain intact and their digested peptides have preserved allergenic capacity highlighting their important roles in allergic reactions to peanut.",
publisher = "Blackwell Publishing Ltd",
journal = "Clinical and Experimental Allergy",
title = "Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides",
pages = "740-731",
number = "6",
volume = "48",
doi = "10.1111/cea.13113"
}

Prodić, I., Stanić-Vučinić, D., Apostolović, D., Mihailović, J., Radibratović, M., Radosavljević, J., Burazer, L., Milcić, M., Smiljanić, K., van Hage, M.,& Ćirković-Veličković, T.. (2018). Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides. in Clinical and Experimental Allergy
Blackwell Publishing Ltd., 48(6), 731-740.
https://doi.org/10.1111/cea.13113

Prodić I, Stanić-Vučinić D, Apostolović D, Mihailović J, Radibratović M, Radosavljević J, Burazer L, Milcić M, Smiljanić K, van Hage M, Ćirković-Veličković T. Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides. in Clinical and Experimental Allergy. 2018;48(6):731-740.
doi:10.1111/cea.13113 .

Prodić, Ivana, Stanić-Vučinić, Dragana, Apostolović, Danijela, Mihailović, Jelena, Radibratović, M., Radosavljević, Jelena, Burazer, Lidija, Milcić, M., Smiljanić, Katarina, van Hage, Marianne, Ćirković-Veličković, Tanja, "Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides" in Clinical and Experimental Allergy, 48, no. 6 (2018):731-740,
https://doi.org/10.1111/cea.13113 . .

TY  - JOUR
AU  - Prodić, Ivana
AU  - Stanić-Vučinić, Dragana
AU  - Apostolović, Danijela
AU  - Mihailović, Jelena
AU  - Radibratović, M.
AU  - Radosavljević, Jelena
AU  - Burazer, Lidija
AU  - Milcić, M.
AU  - Smiljanić, Katarina
AU  - van Hage, Marianne
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/514
AB  - Background: Most food allergens sensitizing via the gastrointestinal tract are stable proteins that are resistant to pepsin digestion, in particular major peanut allergens, Ara h 2 and Ara h 6. Survival of their large fragments is essential for sensitizing capacity. However, the immunoreactive proteins/peptides to which the immune system of the gastrointestinal tract is exposed during digestion of peanut proteins are unknown. Particularly, the IgE reactivity of short digestion-resistant peptides (SDRPs;  lt 10 kDa) released by gastric digestion under standardized and physiologically relevant in vitro conditions has not been investigated. Objective: The aim of this study was to investigate and identify digestion products of major peanut allergens and in particular to examine IgE reactivity of SDRPs released by pepsin digestion of whole peanut grains. Methods: Two-dimensional gel-based proteomics and shotgun peptidomics, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, including far ultraviolet-circular dichroism spectroscopy were used to identify and characterize peanut digesta. Results: Ara h 2 and Ara h 6 remained mostly intact, and SDRPs from Ara h 2 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h 3 exhibited sequential digestion into a series of digestion-resistant peptides with preserved allergenic capacity. A high number of identified SDRPs from Ara h 1, Ara h 2 and Ara h 3 were part of short continuous epitope sequences and possessed substantial allergenic potential. Conclusion and Clinical Relevance: Peanut grain digestion by oral and gastric phase enzymes generates mixture of products, where the major peanut allergens remain intact and their digested peptides have preserved allergenic capacity highlighting their important roles in allergic reactions to peanut.
PB  - Blackwell Publishing Ltd
T2  - Clinical and Experimental Allergy
T1  - Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides
EP  - 740
IS  - 6
SP  - 731
VL  - 48
DO  - 10.1111/cea.13113
ER  - 

@article{
author = "Prodić, Ivana and Stanić-Vučinić, Dragana and Apostolović, Danijela and Mihailović, Jelena and Radibratović, M. and Radosavljević, Jelena and Burazer, Lidija and Milcić, M. and Smiljanić, Katarina and van Hage, Marianne and Ćirković-Veličković, Tanja",
year = "2018",
abstract = "Background: Most food allergens sensitizing via the gastrointestinal tract are stable proteins that are resistant to pepsin digestion, in particular major peanut allergens, Ara h 2 and Ara h 6. Survival of their large fragments is essential for sensitizing capacity. However, the immunoreactive proteins/peptides to which the immune system of the gastrointestinal tract is exposed during digestion of peanut proteins are unknown. Particularly, the IgE reactivity of short digestion-resistant peptides (SDRPs;  lt 10 kDa) released by gastric digestion under standardized and physiologically relevant in vitro conditions has not been investigated. Objective: The aim of this study was to investigate and identify digestion products of major peanut allergens and in particular to examine IgE reactivity of SDRPs released by pepsin digestion of whole peanut grains. Methods: Two-dimensional gel-based proteomics and shotgun peptidomics, immunoblotting with allergen-specific antibodies from peanut-sensitized patients, enzyme-linked immunosorbent inhibition assay and ImmunoCAP tests, including far ultraviolet-circular dichroism spectroscopy were used to identify and characterize peanut digesta. Results: Ara h 2 and Ara h 6 remained mostly intact, and SDRPs from Ara h 2 were more potent in inhibiting IgE binding than Ara h 1 and Ara 3. Ara h 1 and Ara h 3 exhibited sequential digestion into a series of digestion-resistant peptides with preserved allergenic capacity. A high number of identified SDRPs from Ara h 1, Ara h 2 and Ara h 3 were part of short continuous epitope sequences and possessed substantial allergenic potential. Conclusion and Clinical Relevance: Peanut grain digestion by oral and gastric phase enzymes generates mixture of products, where the major peanut allergens remain intact and their digested peptides have preserved allergenic capacity highlighting their important roles in allergic reactions to peanut.",
publisher = "Blackwell Publishing Ltd",
journal = "Clinical and Experimental Allergy",
title = "Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides",
pages = "740-731",
number = "6",
volume = "48",
doi = "10.1111/cea.13113"
}

Prodić, I., Stanić-Vučinić, D., Apostolović, D., Mihailović, J., Radibratović, M., Radosavljević, J., Burazer, L., Milcić, M., Smiljanić, K., van Hage, M.,& Ćirković-Veličković, T.. (2018). Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides. in Clinical and Experimental Allergy
Blackwell Publishing Ltd., 48(6), 731-740.
https://doi.org/10.1111/cea.13113

Prodić I, Stanić-Vučinić D, Apostolović D, Mihailović J, Radibratović M, Radosavljević J, Burazer L, Milcić M, Smiljanić K, van Hage M, Ćirković-Veličković T. Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides. in Clinical and Experimental Allergy. 2018;48(6):731-740.
doi:10.1111/cea.13113 .

Prodić, Ivana, Stanić-Vučinić, Dragana, Apostolović, Danijela, Mihailović, Jelena, Radibratović, M., Radosavljević, Jelena, Burazer, Lidija, Milcić, M., Smiljanić, Katarina, van Hage, Marianne, Ćirković-Veličković, Tanja, "Influence of peanut matrix on stability of allergens in gastric-simulated digesta: 2S albumins are main contributors to the IgE reactivity of short digestion-resistant peptides" in Clinical and Experimental Allergy, 48, no. 6 (2018):731-740,
https://doi.org/10.1111/cea.13113 . .

TY  - CONF
AU  - Radosavljević, Jelena
AU  - Apostolović, Danijela
AU  - Mihailović, Jelena
AU  - Atanasković-Marković, Marina
AU  - Burazer, Lidija
AU  - van Hage, Marianne
AU  - Ćirković-Veličković, Tanja
PY  - 2018
UR  - http://intor.torlakinstitut.com/handle/123456789/504
PB  - Wiley, Hoboken
C3  - FEBS Open Bio
T1  - Digestomics of cow's milk: casein-derived digestion-resistant peptides aggregate into functional complexes
EP  - 257
SP  - 257
VL  - 8
UR  - https://hdl.handle.net/21.15107/rcub_intor_504
ER  - 

@conference{
author = "Radosavljević, Jelena and Apostolović, Danijela and Mihailović, Jelena and Atanasković-Marković, Marina and Burazer, Lidija and van Hage, Marianne and Ćirković-Veličković, Tanja",
year = "2018",
publisher = "Wiley, Hoboken",
journal = "FEBS Open Bio",
title = "Digestomics of cow's milk: casein-derived digestion-resistant peptides aggregate into functional complexes",
pages = "257-257",
volume = "8",
url = "https://hdl.handle.net/21.15107/rcub_intor_504"
}

Radosavljević, J., Apostolović, D., Mihailović, J., Atanasković-Marković, M., Burazer, L., van Hage, M.,& Ćirković-Veličković, T.. (2018). Digestomics of cow's milk: casein-derived digestion-resistant peptides aggregate into functional complexes. in FEBS Open Bio
Wiley, Hoboken., 8, 257-257.
https://hdl.handle.net/21.15107/rcub_intor_504

Radosavljević J, Apostolović D, Mihailović J, Atanasković-Marković M, Burazer L, van Hage M, Ćirković-Veličković T. Digestomics of cow's milk: casein-derived digestion-resistant peptides aggregate into functional complexes. in FEBS Open Bio. 2018;8:257-257.
https://hdl.handle.net/21.15107/rcub_intor_504 .

Radosavljević, Jelena, Apostolović, Danijela, Mihailović, Jelena, Atanasković-Marković, Marina, Burazer, Lidija, van Hage, Marianne, Ćirković-Veličković, Tanja, "Digestomics of cow's milk: casein-derived digestion-resistant peptides aggregate into functional complexes" in FEBS Open Bio, 8 (2018):257-257,
https://hdl.handle.net/21.15107/rcub_intor_504 .