TY  - JOUR
AU  - Pavlović, Marija
AU  - Margetić, Aleksandra
AU  - Leonardi, Adrijana
AU  - Križaj, Igor
AU  - Kojić, Milan
AU  - Vujčić, Zoran
AU  - Šokarda Slavić, Marinela
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/863
AB  - This study focuses on the isolation, purification, and characterisation of endo-polygalacturonase II from Aspergillus tubingensis FAT43, particularly emphasising its potential applications in the fruit juice industry. A comprehensive screening test revealed the temporal dynamics of endo-polygalacturonase production during a 96-hour fermentation process. The purification process, involving ammonium sulfate and ethanol precipitation followed by ion-exchange chromatography, resulted in a 3.3-fold purification of PG II with a yield of 16% and a specific activity of 6001.67 U mg−1. Molecular analysis confirmed the identity of PG II, its gene (pgaII), and a high degree of sequence identity with Aspergillus tubingensis in the SWISS-PROT database. The optimal pH for PG II activity was 3.5–4.5, with robust stability across a broad pH spectrum (3–7). The enzyme exhibited optimal temperature activity at 45 °C, with a retention of 90% activity at 50 °C. The calculated activation energy for PG II was 62.1 kJ mol−1, indicating good stability. Inactivation kinetics revealed a half-life of 13.7 h at 40 °C, 5.4 h at 50 °C, and 0.85 h at 60 °C, with an activation energy of denaturation of 32.8 kJ mol−1. Compared to literature-reported PGs, PG II from A. tubingensis FAT43 demonstrated superior thermal stability. Hydrolysis experiments on different pectins revealed the highest specificity for non-methylated substrates (polygalacturonic acid). In fruit juice processing, PG II significantly increased juice yield and clarity, with the highest impact observed in strawberry juice. Antioxidant activity assays indicated enhanced antioxidant potential in enzyme-treated juices, especially strawberry, quince, and apple juices. The study highlights PG II's potential as an industrially valuable enzyme for fruit juice processing, offering improved thermostability and versatility across various fruit types.
PB  - Royal Society of Chemistry
T2  - Food & Function
T1  - Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential
DO  - 10.1039/D3FO05297D
ER  - 

@article{
author = "Pavlović, Marija and Margetić, Aleksandra and Leonardi, Adrijana and Križaj, Igor and Kojić, Milan and Vujčić, Zoran and Šokarda Slavić, Marinela",
year = "2024",
abstract = "This study focuses on the isolation, purification, and characterisation of endo-polygalacturonase II from Aspergillus tubingensis FAT43, particularly emphasising its potential applications in the fruit juice industry. A comprehensive screening test revealed the temporal dynamics of endo-polygalacturonase production during a 96-hour fermentation process. The purification process, involving ammonium sulfate and ethanol precipitation followed by ion-exchange chromatography, resulted in a 3.3-fold purification of PG II with a yield of 16% and a specific activity of 6001.67 U mg−1. Molecular analysis confirmed the identity of PG II, its gene (pgaII), and a high degree of sequence identity with Aspergillus tubingensis in the SWISS-PROT database. The optimal pH for PG II activity was 3.5–4.5, with robust stability across a broad pH spectrum (3–7). The enzyme exhibited optimal temperature activity at 45 °C, with a retention of 90% activity at 50 °C. The calculated activation energy for PG II was 62.1 kJ mol−1, indicating good stability. Inactivation kinetics revealed a half-life of 13.7 h at 40 °C, 5.4 h at 50 °C, and 0.85 h at 60 °C, with an activation energy of denaturation of 32.8 kJ mol−1. Compared to literature-reported PGs, PG II from A. tubingensis FAT43 demonstrated superior thermal stability. Hydrolysis experiments on different pectins revealed the highest specificity for non-methylated substrates (polygalacturonic acid). In fruit juice processing, PG II significantly increased juice yield and clarity, with the highest impact observed in strawberry juice. Antioxidant activity assays indicated enhanced antioxidant potential in enzyme-treated juices, especially strawberry, quince, and apple juices. The study highlights PG II's potential as an industrially valuable enzyme for fruit juice processing, offering improved thermostability and versatility across various fruit types.",
publisher = "Royal Society of Chemistry",
journal = "Food & Function",
title = "Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential",
doi = "10.1039/D3FO05297D"
}

Pavlović, M., Margetić, A., Leonardi, A., Križaj, I., Kojić, M., Vujčić, Z.,& Šokarda Slavić, M.. (2024). Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential. in Food & Function
Royal Society of Chemistry..
https://doi.org/10.1039/D3FO05297D

Pavlović M, Margetić A, Leonardi A, Križaj I, Kojić M, Vujčić Z, Šokarda Slavić M. Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential. in Food & Function. 2024;.
doi:10.1039/D3FO05297D .

Pavlović, Marija, Margetić, Aleksandra, Leonardi, Adrijana, Križaj, Igor, Kojić, Milan, Vujčić, Zoran, Šokarda Slavić, Marinela, "Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential" in Food & Function (2024),
https://doi.org/10.1039/D3FO05297D . .

TY  - DATA
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/859
AB  - S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.
IS  - 1
VL  - 25
UR  - https://hdl.handle.net/21.15107/rcub_intor_859
ER  - 

@misc{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "S1.1. Checking of N protein purity Recombinant N protein purity was checked after imidazole removal and buffer exchange by SDS PAGE (Figure 6.). For comparison, commercial high-purity HSA was also analyzed. S1.2. Identification of N protein Tandem mass spectrometry identification of proteins in an in-gel digested band of N protein (Figure S1, lane 3), confirmed the identity of N protein with high scores and peptide coverage (Fig. S2.). S2. Purification of polyclonal antibodies from mice and rabbit sera For the development of an ELISA test specific for the detection of SARS-CoV-2 N protein, recombinantly produced N protein was used for the immunization of mice and rabbits. Sera obtained from rabbits and mice were then tested for titer and specificity (Figure S3 and Figure 1). To determine the titer of polyclonal sera required to detect N protein in samples, we use wells coated with N protein and serial dilution of sera pools from different animals. After multiple washing steps, we detected the binding of rabbit and mice antibodies using secondary biotinylated antibodies and streptavidin-alkaline phosphatase chimaera or secondary antibodies with previously coupled alkaline phosphatase, where the amount of enzymes’ substrate converted to the product was measured as an increase in absorbance at 405 nm. As shown in Figure S3A, unpurified sera pools from both animals showed very high titers and expected logarithmic decrease of signal with dilution. Based on the obtained data titer for unpurified sera was determined to be X. The same trend was observed for pools purified using AS precipitation and rabbit sera purified using protein A affinity chromatography (Figure S3B and S3C). As shown in Figure S3D, clear bands from antibodies could be observed in both full and purified samples. Western blot analysis showed only one protein band on mass around 40 kDa, a Accession number / Protein Name Score Coverage (%) Unique peptides P0DTC9|NCAP_SARS2 Nucleoprotein OS=Severe acute respiratory syndrome coronavirus 2, 46 kDa 504.9 74.22 183 mass of purified N protein suggesting that the obtained sera is highly specific for N protein (Figure 2). Section S3 Diagnostic validationS3.1. Stabilization of capture antibodies Pre-coated ELISA plates were prepared for usage in clinical practice. To ensure the preservation of the biofunctionality of the surface-bound capture antibodies, the commonly used stabilizing excipient, 3% sucrose with 10% glycerol in MilliQ water was used. The plates were incubated with 300 μL per well of a stabilizing agent for 1 hour at room temperature. After an hour of incubation, the solution was carefully aspirated from each well. The plate was then blotted against clear paper towels to remove any remaining liquid, and the plates were allowed to air dry for 3 hours at RT. Dried plates were wrapped in parafilm and stored at 4 °C for later use. To remove the stabilizing agent coating, wells were washed with slightly acidic distilled water (pH of 6) three times, leaving the plate prepared for subsequent assay steps. Section S4. Characterization of N protein by HRMS S4.1. SDS PAGE and in-gel digestion Characterization of the produced recombinant N protein was done by HRMS after its in-gel digestion. A total of 10 μg of purified protein(s) were loaded in a 0.5 cm wide well and after SDSPAGE gel was stained with Coomassie Brilliant Blue R-250 (CBB). Protein gel bands were washed, reduced with dithiothreitol, and alkylated with iodoacetamide, followed by in-gel trypsin digestion1 (Shevchenko et al. 2006) with some minor modifications. The amount of trypsin was leveled to a trypsin/sample ratio of 1:30 (w/w). The final concentration of MS-grade trypsin (diluted in 25 mM ammonium bicarbonate buffer) was 1 ng/μL. Sample clean-up was performed using zip tips HyperSep C18 (Thermo Fisher Scientific Inc., Bremen, Germany). S5.1 Immunization of rabbits and mice Mice immunization Swiss Webster mice (n=10) were immunized subcutaneously with N protein formulated with Complete Freund`s adjuvant (CFA; 1st dose, 100 μg N protein / dose) or Incomplete Freund`s adjuvant (IFA; 2nd and 3rd doses, 50 μg N protein / dose) in three-week intervals. Mice were housed in small groups of up to six animals and had access to commercial mice food and water ad libitum. N protein solution (500ug/ml in PBS) was sterilized by filtering through 0.22 um filters. Sterile N protein solution was mixed with CFA (Sigma, Cat. No. F5881) at ratio 1:1 (v/v) under aseptic conditions. In total 400 ul of N protein-CFA emulsion (N protein final concentration 250ug/ml) was applied per immunization per mouse. Initial immunization was done by injection of N protein in CFA given subcutaneously (SC) in four sites (thigh pocket, base of tail, and mediastinum) with a 100 ul using 23-25 gauge needle. In total 100 ug of N protein was applied per mouse (25 ug per site). Subsequent immunizations with booster doses were done in the same way, but using IFA (Sigma, Cat. No. F5506) instead of CFA and N protein final concentration was 125 ug/ml. . In total 50 ug of N protein was applied per mouse (12.5 ug per site). Immunizations were done every three weeks. Mice immunization scheme: 1. day 0 – N protein in PBS: CFA = 1:1 (v/v); N protein final concentration was 250 μg/mL; 400 μL per mice (4x100 μL), e.g. 100 μg per mice 2. day 21 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μL per mice (4x100 μL), e.g. 50 μg per mice 3. day 42 - N protein in PBS: IFA = 1:1 (v/v); N protein final concentration was 125 μg/mL; 400 μl per mice (4x100 μL), e.g. 50 μg per mice First bleeding was performed two weeks after the 3rd dose, and then in intervals not shorter than two weeks. The sera obtained after the first bleeding was tested for the production of specific anti-N protein antibodies.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.",
number = "1",
volume = "25",
url = "https://hdl.handle.net/21.15107/rcub_intor_859"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences
MDPI., 25(1).
https://hdl.handle.net/21.15107/rcub_intor_859

Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333.. in International Journal of Molecular Sciences. 2024;25(1).
https://hdl.handle.net/21.15107/rcub_intor_859 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Supplementary information for the article:       Mladenovic Stokanic, M.; Simovic, A.; Jovanovic, V.; Radomirovic, M.; Udovicki, B.; Krstic Ristivojevic, M.; Djukic, T.; Vasovic, T.; Acimovic, J.; Sabljic, L.; Lukic, I.; Kovacevic, A.; Cujic, D.; Gnjatovic, M.; Smiljanic, K.; Stojadinovic, M.; Radosavljevic, J.; Stanic-Vucinic, D.; Stojanovic, M.; Rajkovic, A.; Cirkovic Velickovic, T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. International Journal of Molecular Sciences 2024, 25 (1), 333. https://doi.org/10.3390/ijms25010333." in International Journal of Molecular Sciences, 25, no. 1 (2024),
https://hdl.handle.net/21.15107/rcub_intor_859 .

TY  - JOUR
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana
AU  - Udovički, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Djukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujic, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirkovic Veličković, Tanja
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/858
AB  - In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species
IS  - 1
SP  - 333
VL  - 25
DO  - 10.3390/ijms25010333
ER  - 

@article{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna and Radomirović, Mirjana and Udovički, Božidar and Krstić Ristivojević, Maja and Djukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujic, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirkovic Veličković, Tanja",
year = "2024",
abstract = "In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species",
number = "1",
pages = "333",
volume = "25",
doi = "10.3390/ijms25010333"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V., Radomirović, M., Udovički, B., Krstić Ristivojević, M., Djukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujic, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirkovic Veličković, T.. (2024). Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences
MDPI., 25(1), 333.
https://doi.org/10.3390/ijms25010333

Mladenović Stokanić M, Simović A, Jovanović V, Radomirović M, Udovički B, Krstić Ristivojević M, Djukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujic D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirkovic Veličković T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences. 2024;25(1):333.
doi:10.3390/ijms25010333 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna, Radomirović, Mirjana, Udovički, Božidar, Krstić Ristivojević, Maja, Djukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujic, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirkovic Veličković, Tanja, "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species" in International Journal of Molecular Sciences, 25, no. 1 (2024):333,
https://doi.org/10.3390/ijms25010333 . .

TY  - JOUR
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Trifunovic, Sara
AU  - Krstić Ristivojević, Maja
AU  - Aćimović, Milica
AU  - Stanković Jeremić, Jovana
AU  - Lončar, Biljana
AU  - Tešević, Vele
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/857
AB  - As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.
PB  - MDPI
T2  - Antioxidants
T2  - Antioxidants
T1  - Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts
IS  - 11
SP  - 1988
VL  - 12
DO  - 10.3390/antiox12111988
ER  - 

@article{
author = "Smiljanić, Katarina and Prodić, Ivana and Trifunovic, Sara and Krstić Ristivojević, Maja and Aćimović, Milica and Stanković Jeremić, Jovana and Lončar, Biljana and Tešević, Vele",
year = "2023",
abstract = "As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.",
publisher = "MDPI",
journal = "Antioxidants, Antioxidants",
title = "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts",
number = "11",
pages = "1988",
volume = "12",
doi = "10.3390/antiox12111988"
}

Smiljanić, K., Prodić, I., Trifunovic, S., Krstić Ristivojević, M., Aćimović, M., Stanković Jeremić, J., Lončar, B.,& Tešević, V.. (2023). Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants
MDPI., 12(11), 1988.
https://doi.org/10.3390/antiox12111988

Smiljanić K, Prodić I, Trifunovic S, Krstić Ristivojević M, Aćimović M, Stanković Jeremić J, Lončar B, Tešević V. Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants. 2023;12(11):1988.
doi:10.3390/antiox12111988 .

Smiljanić, Katarina, Prodić, Ivana, Trifunovic, Sara, Krstić Ristivojević, Maja, Aćimović, Milica, Stanković Jeremić, Jovana, Lončar, Biljana, Tešević, Vele, "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts" in Antioxidants, 12, no. 11 (2023):1988,
https://doi.org/10.3390/antiox12111988 . .

TY  - JOUR
AU  - Prodić, Ivana
AU  - Krstić-Ristivojević, Maja
AU  - Smiljanić, Katarina
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/756
AB  - Thermally processed peanuts are ideal plant models for studying the relationship between allergenicity and antioxidant capacity of protein-rich foods, besides lipids, carbohydrates and phytochemicals. Peanut is highly praised in the human diet; however, it is rich in allergens (>75% of total proteins). One-third of peanut allergens belong to the products of genes responsible for the defence of plants against stress conditions. The proximate composition of major peanut macromolecules and polyphenols is reviewed, focusing on the identity and relative abundance of all peanut proteins derived from recent proteomic studies. The importance of thermal processing, gastrointestinal digestion (performed by INFOGEST protocol) and their influence on allergenicity and antioxidant properties of protein-rich plant food matrices is elaborated. Antioxidant properties of bioactive peptides from nuts were also considered. Moreover, there are no studies dealing simultaneously with the antioxidant and allergenic properties of protein- and polyphenol-rich foods, considering all the molecules that can significantly contribute to the antioxidant capacity during and after gastrointestinal digestion. In summary, proteins and carbohydrates are underappreciated sources of antioxidant power released during the gastrointestinal digestion of protein-rich plant foods, and it is crucial to decipher their antioxidant contribution in addition to polyphenols and vitamins before and after gastrointestinal digestion.
PB  - MDPI
T2  - Antioxidants
T1  - Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies
IS  - 4
SP  - 886
VL  - 12
DO  - 10.3390/antiox12040886
ER  - 

@article{
author = "Prodić, Ivana and Krstić-Ristivojević, Maja and Smiljanić, Katarina",
year = "2023",
abstract = "Thermally processed peanuts are ideal plant models for studying the relationship between allergenicity and antioxidant capacity of protein-rich foods, besides lipids, carbohydrates and phytochemicals. Peanut is highly praised in the human diet; however, it is rich in allergens (>75% of total proteins). One-third of peanut allergens belong to the products of genes responsible for the defence of plants against stress conditions. The proximate composition of major peanut macromolecules and polyphenols is reviewed, focusing on the identity and relative abundance of all peanut proteins derived from recent proteomic studies. The importance of thermal processing, gastrointestinal digestion (performed by INFOGEST protocol) and their influence on allergenicity and antioxidant properties of protein-rich plant food matrices is elaborated. Antioxidant properties of bioactive peptides from nuts were also considered. Moreover, there are no studies dealing simultaneously with the antioxidant and allergenic properties of protein- and polyphenol-rich foods, considering all the molecules that can significantly contribute to the antioxidant capacity during and after gastrointestinal digestion. In summary, proteins and carbohydrates are underappreciated sources of antioxidant power released during the gastrointestinal digestion of protein-rich plant foods, and it is crucial to decipher their antioxidant contribution in addition to polyphenols and vitamins before and after gastrointestinal digestion.",
publisher = "MDPI",
journal = "Antioxidants",
title = "Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies",
number = "4",
pages = "886",
volume = "12",
doi = "10.3390/antiox12040886"
}

Prodić, I., Krstić-Ristivojević, M.,& Smiljanić, K.. (2023). Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies. in Antioxidants
MDPI., 12(4), 886.
https://doi.org/10.3390/antiox12040886

Prodić I, Krstić-Ristivojević M, Smiljanić K. Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies. in Antioxidants. 2023;12(4):886.
doi:10.3390/antiox12040886 .

Prodić, Ivana, Krstić-Ristivojević, Maja, Smiljanić, Katarina, "Antioxidant Properties of Protein-Rich Plant Foods in Gastrointestinal Digestion-Peanuts as Our Antioxidant Friend or Foe in Allergies" in Antioxidants, 12, no. 4 (2023):886,
https://doi.org/10.3390/antiox12040886 . .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/788
AB  - Food allergies have increased dramatically in the last decade, especially in developedcountries. Food tolerance requires strict maintenance of a specific microbial portfolio inthe gastrointestinal tract, as changes in the gut microbiome can lead to its disruption,which in turn causes inflammation and pathogenic gut conditions leading to thedevelopment of food allergies. Any environmental factors that lead to a disturbanceand/or malfunction of the gastrointestinal tract and digestive performance favor thedevelopment of food allergies.Based on that, what do we know about the role of increasing anthropogenic chemicals,including emerging ones, resulting from the new global situation?There is awareness that their effects are multifaceted, e.g., chemicals affect the growth ofplants and animals and thus the quality of the food produced. In addition, chemicals affectour food during its production and processing, but also affect our body andgastrointestinal tract. It is time to fill the knowledge gaps and understand how theseinteractions between environmental triggers such as industrial and traffic pollution,transition and heavy metals, pesticides, chemtrails, etc., affect food allergens and theirallergenicity, adjuvant effects, and the increasing prevalence of food allergies.Some improvements in this area are already being made through advances in ‘omics’technologies (i.e., proteomics, genomics, metabolomics) and systems biology approachesthat will hopefully provide a scientific understanding of the relationship betweenincreasing food allergies and the increasingly present wide variety of anthropogenicchemicals in our environment.
PB  - Udruženje za preventivnu pedijatriju Srbije
C3  - Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.
T1  - Food allergies on the rise: the role of anthropogenic chemicals
EP  - 27
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_intor_788
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana",
year = "2023",
abstract = "Food allergies have increased dramatically in the last decade, especially in developedcountries. Food tolerance requires strict maintenance of a specific microbial portfolio inthe gastrointestinal tract, as changes in the gut microbiome can lead to its disruption,which in turn causes inflammation and pathogenic gut conditions leading to thedevelopment of food allergies. Any environmental factors that lead to a disturbanceand/or malfunction of the gastrointestinal tract and digestive performance favor thedevelopment of food allergies.Based on that, what do we know about the role of increasing anthropogenic chemicals,including emerging ones, resulting from the new global situation?There is awareness that their effects are multifaceted, e.g., chemicals affect the growth ofplants and animals and thus the quality of the food produced. In addition, chemicals affectour food during its production and processing, but also affect our body andgastrointestinal tract. It is time to fill the knowledge gaps and understand how theseinteractions between environmental triggers such as industrial and traffic pollution,transition and heavy metals, pesticides, chemtrails, etc., affect food allergens and theirallergenicity, adjuvant effects, and the increasing prevalence of food allergies.Some improvements in this area are already being made through advances in ‘omics’technologies (i.e., proteomics, genomics, metabolomics) and systems biology approachesthat will hopefully provide a scientific understanding of the relationship betweenincreasing food allergies and the increasingly present wide variety of anthropogenicchemicals in our environment.",
publisher = "Udruženje za preventivnu pedijatriju Srbije",
journal = "Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.",
title = "Food allergies on the rise: the role of anthropogenic chemicals",
pages = "27-27",
url = "https://hdl.handle.net/21.15107/rcub_intor_788"
}

Smiljanić, K.,& Prodić, I.. (2023). Food allergies on the rise: the role of anthropogenic chemicals. in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.
Udruženje za preventivnu pedijatriju Srbije., 27-27.
https://hdl.handle.net/21.15107/rcub_intor_788

Smiljanić K, Prodić I. Food allergies on the rise: the role of anthropogenic chemicals. in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.. 2023;:27-27.
https://hdl.handle.net/21.15107/rcub_intor_788 .

Smiljanić, Katarina, Prodić, Ivana, "Food allergies on the rise: the role of anthropogenic chemicals" in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023. (2023):27-27,
https://hdl.handle.net/21.15107/rcub_intor_788 .

TY  - CONF
AU  - Protić-Rosić, Isidora
AU  - Lopandić, Zorana
AU  - Popović, Dragan
AU  - Blagojević, Gordan
AU  - Gavrović-Jankulović, Marija
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/714
AB  - Allergen immunotherapy (AIT) is currently the only disease-modifying treatment for
allergies. Pre-clinical models for the evaluation of novel therapeutics are crucial for
ensuring their efficacy and safety. While cell culture models are cost-effective and
efficient, they cannot fully replicate the cellular interactions in vivo. Therefore, it is
essential to use more sophisticated model systems, such as co-cultures, to assess the
potential of new therapeutics more accurately. Immunomodulatory protein banana lectin
(BLwt) is an attractive candidate for adjuvant in AIT. Its mutant BLH84T was developed to
reduce its potential mitogenicity. The aim of this study was the development of the coculture model system for testing the immunomodulatory effect of chimeras composed of
the major birch pollen allergen (Bv1a) and BLwt (Bv1a-BLwt, Cwt), the hypoallergenic
isoform of Bv1a (Bv1l) and BLH84T (Bv1l-BLH84T, C1 and BLH84T-Bv1l, C2). Chimeric
structures were designed in silico, fully minimized, and relaxed without van der Waals
atomic clashes. Afterward, proteins were successfully expressed in Escherichia coli and
purified by IMAC yielding around 0.4 mg per 1L of expression medium. The IgE binding
capacity was assessed using ELISA inhibition with birch pollen allergic patients’ sera.
Caco-2 intestinal epithelial cells and THP-1 differentiated macrophages were used for the
co-culture model system development. After protein application on the apical side of the
co-culture, the integrity of the epithelial monolayer was not disturbed. The
immunomodulatory potential of antigens was tested by measuring the gene expression
levels for pro- and anti-inflammatory cytokines in both cell lines from co-culture. The
obtained results indicate that the best anti-inflammatory response was favored after
treatment with Cwt. Additionally, to further confirm the immunomodulatory effect of the
recombinant chimeras, PBMCs obtained from individuals allergic to birch pollen were
employed and treated with recombinant proteins. Only after treatment with Cwt, PBMCs
secreted the anti-inflammatory cytokine IL-10. Obtained results suggest that Cwt chimera
could have a therapeutic effect in AIT in birch pollen allergy.
PB  - Faculty of Chemistry
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference International scientific meeting September 21-23, 2023, Belgrade, Serbia “Biochemistry in Biotechnology”
T1  - Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system
EP  - 72
SP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_intor_714
ER  - 

@conference{
author = "Protić-Rosić, Isidora and Lopandić, Zorana and Popović, Dragan and Blagojević, Gordan and Gavrović-Jankulović, Marija",
year = "2023",
abstract = "Allergen immunotherapy (AIT) is currently the only disease-modifying treatment for
allergies. Pre-clinical models for the evaluation of novel therapeutics are crucial for
ensuring their efficacy and safety. While cell culture models are cost-effective and
efficient, they cannot fully replicate the cellular interactions in vivo. Therefore, it is
essential to use more sophisticated model systems, such as co-cultures, to assess the
potential of new therapeutics more accurately. Immunomodulatory protein banana lectin
(BLwt) is an attractive candidate for adjuvant in AIT. Its mutant BLH84T was developed to
reduce its potential mitogenicity. The aim of this study was the development of the coculture model system for testing the immunomodulatory effect of chimeras composed of
the major birch pollen allergen (Bv1a) and BLwt (Bv1a-BLwt, Cwt), the hypoallergenic
isoform of Bv1a (Bv1l) and BLH84T (Bv1l-BLH84T, C1 and BLH84T-Bv1l, C2). Chimeric
structures were designed in silico, fully minimized, and relaxed without van der Waals
atomic clashes. Afterward, proteins were successfully expressed in Escherichia coli and
purified by IMAC yielding around 0.4 mg per 1L of expression medium. The IgE binding
capacity was assessed using ELISA inhibition with birch pollen allergic patients’ sera.
Caco-2 intestinal epithelial cells and THP-1 differentiated macrophages were used for the
co-culture model system development. After protein application on the apical side of the
co-culture, the integrity of the epithelial monolayer was not disturbed. The
immunomodulatory potential of antigens was tested by measuring the gene expression
levels for pro- and anti-inflammatory cytokines in both cell lines from co-culture. The
obtained results indicate that the best anti-inflammatory response was favored after
treatment with Cwt. Additionally, to further confirm the immunomodulatory effect of the
recombinant chimeras, PBMCs obtained from individuals allergic to birch pollen were
employed and treated with recombinant proteins. Only after treatment with Cwt, PBMCs
secreted the anti-inflammatory cytokine IL-10. Obtained results suggest that Cwt chimera
could have a therapeutic effect in AIT in birch pollen allergy.",
publisher = "Faculty of Chemistry, Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference International scientific meeting September 21-23, 2023, Belgrade, Serbia “Biochemistry in Biotechnology”",
title = "Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system",
pages = "72-71",
url = "https://hdl.handle.net/21.15107/rcub_intor_714"
}

Protić-Rosić, I., Lopandić, Z., Popović, D., Blagojević, G.,& Gavrović-Jankulović, M.. (2023). Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system. in Serbian Biochemical Society Twelfth Conference International scientific meeting September 21-23, 2023, Belgrade, Serbia “Biochemistry in Biotechnology”
Faculty of Chemistry., 71-72.
https://hdl.handle.net/21.15107/rcub_intor_714

Protić-Rosić I, Lopandić Z, Popović D, Blagojević G, Gavrović-Jankulović M. Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system. in Serbian Biochemical Society Twelfth Conference International scientific meeting September 21-23, 2023, Belgrade, Serbia “Biochemistry in Biotechnology”. 2023;:71-72.
https://hdl.handle.net/21.15107/rcub_intor_714 .

Protić-Rosić, Isidora, Lopandić, Zorana, Popović, Dragan, Blagojević, Gordan, Gavrović-Jankulović, Marija, "Evaluation of the immunomodulatory potential of chimera Bv1a-BLwt and its mutants on the co-culture model system" in Serbian Biochemical Society Twelfth Conference International scientific meeting September 21-23, 2023, Belgrade, Serbia “Biochemistry in Biotechnology” (2023):71-72,
https://hdl.handle.net/21.15107/rcub_intor_714 .

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  - Prodić, Ivana
AU  - Burazer, Lidija
AU  - Đorić, Nataša
AU  - Krstić-Ristivojević, Maja
AU  - Smiljanić, Katarina
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/771
AB  - Background and Aim: Epidemiological studies pointed at the connection betweenpollution (e.g., traffic emissions) and an increased percentage of people suffering fromrespiratory allergies, including the pediatric population. Field studies provided the mostrelevant assessment of the effects of the intensity and variety of urban and industrialcontamination on the structure and allergenic potency of pollen allergens. Therefore, theaim of the present work was to compare allergenic profiles ofDactylis glomerata pollen(DGP) collected in the specific urban and rural areas (Kruševac and suburbs), to assesspollen structures and immunoglobulin E (IgE) reactivity to pollen of school childrenpopulation allergic to grass pollens.
PB  - Udruženje za preventivnu pedijatriju Srbije
C3  - Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.
T1  - Dactylis glomerata grass pollen from urban area releases more sub- pollen particles and has stronger ige response in allergic individuals than rural counterpart
EP  - 58
SP  - 58
UR  - https://hdl.handle.net/21.15107/rcub_intor_771
ER  - 

@conference{
author = "Prodić, Ivana and Burazer, Lidija and Đorić, Nataša and Krstić-Ristivojević, Maja and Smiljanić, Katarina",
year = "2023",
abstract = "Background and Aim: Epidemiological studies pointed at the connection betweenpollution (e.g., traffic emissions) and an increased percentage of people suffering fromrespiratory allergies, including the pediatric population. Field studies provided the mostrelevant assessment of the effects of the intensity and variety of urban and industrialcontamination on the structure and allergenic potency of pollen allergens. Therefore, theaim of the present work was to compare allergenic profiles ofDactylis glomerata pollen(DGP) collected in the specific urban and rural areas (Kruševac and suburbs), to assesspollen structures and immunoglobulin E (IgE) reactivity to pollen of school childrenpopulation allergic to grass pollens.",
publisher = "Udruženje za preventivnu pedijatriju Srbije",
journal = "Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.",
title = "Dactylis glomerata grass pollen from urban area releases more sub- pollen particles and has stronger ige response in allergic individuals than rural counterpart",
pages = "58-58",
url = "https://hdl.handle.net/21.15107/rcub_intor_771"
}

Prodić, I., Burazer, L., Đorić, N., Krstić-Ristivojević, M.,& Smiljanić, K.. (2023). Dactylis glomerata grass pollen from urban area releases more sub- pollen particles and has stronger ige response in allergic individuals than rural counterpart. in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.
Udruženje za preventivnu pedijatriju Srbije., 58-58.
https://hdl.handle.net/21.15107/rcub_intor_771

Prodić I, Burazer L, Đorić N, Krstić-Ristivojević M, Smiljanić K. Dactylis glomerata grass pollen from urban area releases more sub- pollen particles and has stronger ige response in allergic individuals than rural counterpart. in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023.. 2023;:58-58.
https://hdl.handle.net/21.15107/rcub_intor_771 .

Prodić, Ivana, Burazer, Lidija, Đorić, Nataša, Krstić-Ristivojević, Maja, Smiljanić, Katarina, "Dactylis glomerata grass pollen from urban area releases more sub- pollen particles and has stronger ige response in allergic individuals than rural counterpart" in Knjiga apstrakata: Deseti nacionalni kongres Udruženja za preventivnu pedijatriju Srbije (UPPS) sa međunarodnim učešćem, Kopaonik, 21-23. april 2023. (2023):58-58,
https://hdl.handle.net/21.15107/rcub_intor_771 .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Ristović, Marina
AU  - Pavlović, Marija
AU  - Nikolić, Stefan
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/635
AB  - The combination of b-oligosaccharides from enzymatically hydrolysed barley b-glucan has attracted interest recently due to its positive effects on human health. This study aimed to assess the impact of the
endo-b-1,3-1,4-glucanase enzyme from Bacillus subtilis 168 on improving the nutritional and bioactive
properties of barley b-glucan. A new procedure for the isolation of b-glucan was developed, at a lower
temperature (45 °C), enabling purity from starch contamination, without affecting the yield (6 g b-glucan
from 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned into E. coli pQE_Ek enables the high
production and purification (82% yield, 1.8 mg mL 1 and 440 U mg 1
) of an enzyme identical to the
natural one (25.5 kDa). The enzymatic reaction showed high efficiency of b-glucan degradation by recombinant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosylD-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacities
by 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicate
the possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barley bglucan used as functional foods.
PB  - Wiley
T2  - International Journal of Food Science and Technology
T1  - Improvement of nutritional and bioactive properties of barley b-glucan-based food products using Bacillus subtilis 168 endo-b-1,3-1,4-glucanase
DO  - 10.1111/ijfs.16647
ER  - 

@article{
author = "Šokarda Slavić, Marinela and Kojić, Milan and Margetić, Aleksandra and Ristović, Marina and Pavlović, Marija and Nikolić, Stefan and Vujčić, Zoran",
year = "2023",
abstract = "The combination of b-oligosaccharides from enzymatically hydrolysed barley b-glucan has attracted interest recently due to its positive effects on human health. This study aimed to assess the impact of the
endo-b-1,3-1,4-glucanase enzyme from Bacillus subtilis 168 on improving the nutritional and bioactive
properties of barley b-glucan. A new procedure for the isolation of b-glucan was developed, at a lower
temperature (45 °C), enabling purity from starch contamination, without affecting the yield (6 g b-glucan
from 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned into E. coli pQE_Ek enables the high
production and purification (82% yield, 1.8 mg mL 1 and 440 U mg 1
) of an enzyme identical to the
natural one (25.5 kDa). The enzymatic reaction showed high efficiency of b-glucan degradation by recombinant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosylD-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacities
by 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicate
the possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barley bglucan used as functional foods.",
publisher = "Wiley",
journal = "International Journal of Food Science and Technology",
title = "Improvement of nutritional and bioactive properties of barley b-glucan-based food products using Bacillus subtilis 168 endo-b-1,3-1,4-glucanase",
doi = "10.1111/ijfs.16647"
}

Šokarda Slavić, M., Kojić, M., Margetić, A., Ristović, M., Pavlović, M., Nikolić, S.,& Vujčić, Z.. (2023). Improvement of nutritional and bioactive properties of barley b-glucan-based food products using Bacillus subtilis 168 endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology
Wiley..
https://doi.org/10.1111/ijfs.16647

Šokarda Slavić M, Kojić M, Margetić A, Ristović M, Pavlović M, Nikolić S, Vujčić Z. Improvement of nutritional and bioactive properties of barley b-glucan-based food products using Bacillus subtilis 168 endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology. 2023;.
doi:10.1111/ijfs.16647 .

Šokarda Slavić, Marinela, Kojić, Milan, Margetić, Aleksandra, Ristović, Marina, Pavlović, Marija, Nikolić, Stefan, Vujčić, Zoran, "Improvement of nutritional and bioactive properties of barley b-glucan-based food products using Bacillus subtilis 168 endo-b-1,3-1,4-glucanase" in International Journal of Food Science and Technology (2023),
https://doi.org/10.1111/ijfs.16647 . .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Stanisavljević, Nemanja
AU  - Gardijan, Lazar
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://intor.torlakinstitut.com/handle/123456789/634
AB  - α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications
SP  - 126055
VL  - 249
DO  - 10.1016/j.ijbiomac.2023.126055
ER  - 

@article{
author = "Šokarda Slavić, Marinela and Kojić, Milan and Margetić, Aleksandra and Stanisavljević, Nemanja and Gardijan, Lazar and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications",
pages = "126055",
volume = "249",
doi = "10.1016/j.ijbiomac.2023.126055"
}

Šokarda Slavić, M., Kojić, M., Margetić, A., Stanisavljević, N., Gardijan, L., Božić, N.,& Vujčić, Z.. (2023). Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules
Elsevier., 249, 126055.
https://doi.org/10.1016/j.ijbiomac.2023.126055

Šokarda Slavić M, Kojić M, Margetić A, Stanisavljević N, Gardijan L, Božić N, Vujčić Z. Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules. 2023;249:126055.
doi:10.1016/j.ijbiomac.2023.126055 .

Šokarda Slavić, Marinela, Kojić, Milan, Margetić, Aleksandra, Stanisavljević, Nemanja, Gardijan, Lazar, Božić, Nataša, Vujčić, Zoran, "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications" in International Journal of Biological Macromolecules, 249 (2023):126055,
https://doi.org/10.1016/j.ijbiomac.2023.126055 . .

TY  - JOUR
AU  - Lopandić, Zorana
AU  - Dragačević, Luka
AU  - Kosanović, Dejana
AU  - Burazer, Lidija
AU  - Gavrović-Jankulović, Marija
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/787
AB  - In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.
PB  - Elsevier
T2  - Journal of Immunological Methods
T2  - Journal of Immunological MethodsJournal of Immunological Methods
T1  - Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols
SP  - 113382
VL  - 511
DO  - 10.1016/j.jim.2022.113382
ER  - 

@article{
author = "Lopandić, Zorana and Dragačević, Luka and Kosanović, Dejana and Burazer, Lidija and Gavrović-Jankulović, Marija and Minić, Rajna",
year = "2022",
abstract = "In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.",
publisher = "Elsevier",
journal = "Journal of Immunological Methods, Journal of Immunological MethodsJournal of Immunological Methods",
title = "Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols",
pages = "113382",
volume = "511",
doi = "10.1016/j.jim.2022.113382"
}

Lopandić, Z., Dragačević, L., Kosanović, D., Burazer, L., Gavrović-Jankulović, M.,& Minić, R.. (2022). Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols. in Journal of Immunological Methods
Elsevier., 511, 113382.
https://doi.org/10.1016/j.jim.2022.113382

Lopandić Z, Dragačević L, Kosanović D, Burazer L, Gavrović-Jankulović M, Minić R. Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols. in Journal of Immunological Methods. 2022;511:113382.
doi:10.1016/j.jim.2022.113382 .

Lopandić, Zorana, Dragačević, Luka, Kosanović, Dejana, Burazer, Lidija, Gavrović-Jankulović, Marija, Minić, Rajna, "Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols" in Journal of Immunological Methods, 511 (2022):113382,
https://doi.org/10.1016/j.jim.2022.113382 . .

TY  - JOUR
AU  - Trifunović, Sara
AU  - Smiljanić, Katarina
AU  - Sickmann, Albert
AU  - Solari, Fiorella Andrea
AU  - Kolarević, Stoimir
AU  - Divac Rankov, Aleksandra
AU  - Ljujic, Mila
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/781
AB  - Background: Although still considered a safer alternative to classical cigarettes, growing body of work points toharmful effects of electronic cigarettes (e-cigarettes) affecting a range of cellular processes. The biological effect ofe-cigarettes needs to be investigated in more detail considering their widespread use.Methods: In this study, we treated V79 lung fibroblasts with sub-cytotoxic concentration of e-cigarette liquids, withand without nicotine. Mutagenicity was evaluated by HPRT assay, genotoxicity by comet assay and the effect on cel-lular communication by metabolic cooperation assay. Additionally, comprehensive proteome analysis was performedvia high resolution, parallel accumulation serial fragmentation-PASEF mass spectrometry.Results: E-cigarette liquid concentration used in this study showed no mutagenic or genotoxic effect, however itnegatively impacted metabolic cooperation between V79 cells. Both e-cigarette liquids induced significant depletionin total number of proteins and impairment of mitochondrial function in treated cells. The focal adhesion proteinswere upregulated, which is in accordance with the results of metabolic cooperation assay. Increased presence of post-translational modifications (PTMs), including carbonylation and direct oxidative modifications, was observed. Data areavailable via ProteomeXchange with identifier PXD032071.Conclusions: Our study revealed impairment of metabolic cooperation as well as significant proteome and PTMsalterations in V79 cells treated with e-cigarette liquid warranting future studies on e-cigarettes health impact.
PB  - BMC Springer Nature
T2  - Respiratory Research
T1  - Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells
IS  - 191
VL  - 23
DO  - 10.1186/s12931-022-02102-w
ER  - 

@article{
author = "Trifunović, Sara and Smiljanić, Katarina and Sickmann, Albert and Solari, Fiorella Andrea and Kolarević, Stoimir and Divac Rankov, Aleksandra and Ljujic, Mila",
year = "2022",
abstract = "Background: Although still considered a safer alternative to classical cigarettes, growing body of work points toharmful effects of electronic cigarettes (e-cigarettes) affecting a range of cellular processes. The biological effect ofe-cigarettes needs to be investigated in more detail considering their widespread use.Methods: In this study, we treated V79 lung fibroblasts with sub-cytotoxic concentration of e-cigarette liquids, withand without nicotine. Mutagenicity was evaluated by HPRT assay, genotoxicity by comet assay and the effect on cel-lular communication by metabolic cooperation assay. Additionally, comprehensive proteome analysis was performedvia high resolution, parallel accumulation serial fragmentation-PASEF mass spectrometry.Results: E-cigarette liquid concentration used in this study showed no mutagenic or genotoxic effect, however itnegatively impacted metabolic cooperation between V79 cells. Both e-cigarette liquids induced significant depletionin total number of proteins and impairment of mitochondrial function in treated cells. The focal adhesion proteinswere upregulated, which is in accordance with the results of metabolic cooperation assay. Increased presence of post-translational modifications (PTMs), including carbonylation and direct oxidative modifications, was observed. Data areavailable via ProteomeXchange with identifier PXD032071.Conclusions: Our study revealed impairment of metabolic cooperation as well as significant proteome and PTMsalterations in V79 cells treated with e-cigarette liquid warranting future studies on e-cigarettes health impact.",
publisher = "BMC Springer Nature",
journal = "Respiratory Research",
title = "Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells",
number = "191",
volume = "23",
doi = "10.1186/s12931-022-02102-w"
}

Trifunović, S., Smiljanić, K., Sickmann, A., Solari, F. A., Kolarević, S., Divac Rankov, A.,& Ljujic, M.. (2022). Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells. in Respiratory Research
BMC Springer Nature., 23(191).
https://doi.org/10.1186/s12931-022-02102-w

Trifunović S, Smiljanić K, Sickmann A, Solari FA, Kolarević S, Divac Rankov A, Ljujic M. Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells. in Respiratory Research. 2022;23(191).
doi:10.1186/s12931-022-02102-w .

Trifunović, Sara, Smiljanić, Katarina, Sickmann, Albert, Solari, Fiorella Andrea, Kolarević, Stoimir, Divac Rankov, Aleksandra, Ljujic, Mila, "Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells" in Respiratory Research, 23, no. 191 (2022),
https://doi.org/10.1186/s12931-022-02102-w . .

TY  - JOUR
AU  - Lopandic, Zorana
AU  - Dragačević, Luka
AU  - Popovic, Dragan
AU  - Andjelkovic, Uros
AU  - Minić, Rajna
AU  - Gavrovic-Jankulovic, Marija
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/615
AB  - Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms
PB  - MDPI
T2  - Biomolecules
T1  - BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms
IS  - 2
SP  - 180
VL  - 11(2)
VL  - 11
DO  - 10.3390/biom11020180
ER  - 

@article{
author = "Lopandic, Zorana and Dragačević, Luka and Popovic, Dragan and Andjelkovic, Uros and Minić, Rajna and Gavrovic-Jankulovic, Marija",
year = "2021",
abstract = "Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms",
publisher = "MDPI",
journal = "Biomolecules",
title = "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms",
number = "2",
pages = "180",
volume = "11(2), 11",
doi = "10.3390/biom11020180"
}

Lopandic, Z., Dragačević, L., Popovic, D., Andjelkovic, U., Minić, R.,& Gavrovic-Jankulovic, M.. (2021). BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules
MDPI., 11(2)(2), 180.
https://doi.org/10.3390/biom11020180

Lopandic Z, Dragačević L, Popovic D, Andjelkovic U, Minić R, Gavrovic-Jankulovic M. BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules. 2021;11(2)(2):180.
doi:10.3390/biom11020180 .

Lopandic, Zorana, Dragačević, Luka, Popovic, Dragan, Andjelkovic, Uros, Minić, Rajna, Gavrovic-Jankulovic, Marija, "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms" in Biomolecules, 11(2), no. 2 (2021):180,
https://doi.org/10.3390/biom11020180 . .

TY  - JOUR
AU  - Krstić-Ristivojević, Maja
AU  - Apostolović, Danijela
AU  - Smiljanić, Katarina
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/784
AB  - Food hypersensitivity reactions are adverse reactions to harmless dietary substances, whose causes are hidden within derangements of the complex immune machinery of humans and mammals. Until recently, enterocytes were considered as solely absorptive cells providing a physical barrier for unwanted lumen constituents. This review focuses on the enterocytes, which are the hub for innate and adaptive immune reactions. Furthermore, the ambiguous nature of enterocytes is also reflected in the fact that enterocytes can be considered as antigen-presenting cells since they constitutively express major histocompatibility complex (MHC) class II molecules. Taken together, it becomes clear that enterocytes have an immense role in maintaining oral tolerance to foreign antigens. In general, the immune system and its mechanisms underlying food hypersensitivity are still unknown and the involvement of components belonging to other anatomical systems, such as enterocytes, in these mechanisms make their elucidation even more difficult. The findings from studies with animal models provide us with valuable information about allergic mechanisms in the animal world, while on the other hand, these models are used to extrapolate results to the pathological conditions occurring in humans. There is a constant need for studies that deal with this topic and can overcome the glitches related to ethics in working with animals.
PB  - MDPI
T2  - Animals
T1  - Enterocytes in Food Hypersensitivity Reactions
IS  - 9
SP  - 2713
VL  - 11
DO  - 10.3390/ani11092713
ER  - 

@article{
author = "Krstić-Ristivojević, Maja and Apostolović, Danijela and Smiljanić, Katarina",
year = "2021",
abstract = "Food hypersensitivity reactions are adverse reactions to harmless dietary substances, whose causes are hidden within derangements of the complex immune machinery of humans and mammals. Until recently, enterocytes were considered as solely absorptive cells providing a physical barrier for unwanted lumen constituents. This review focuses on the enterocytes, which are the hub for innate and adaptive immune reactions. Furthermore, the ambiguous nature of enterocytes is also reflected in the fact that enterocytes can be considered as antigen-presenting cells since they constitutively express major histocompatibility complex (MHC) class II molecules. Taken together, it becomes clear that enterocytes have an immense role in maintaining oral tolerance to foreign antigens. In general, the immune system and its mechanisms underlying food hypersensitivity are still unknown and the involvement of components belonging to other anatomical systems, such as enterocytes, in these mechanisms make their elucidation even more difficult. The findings from studies with animal models provide us with valuable information about allergic mechanisms in the animal world, while on the other hand, these models are used to extrapolate results to the pathological conditions occurring in humans. There is a constant need for studies that deal with this topic and can overcome the glitches related to ethics in working with animals.",
publisher = "MDPI",
journal = "Animals",
title = "Enterocytes in Food Hypersensitivity Reactions",
number = "9",
pages = "2713",
volume = "11",
doi = "10.3390/ani11092713"
}

Krstić-Ristivojević, M., Apostolović, D.,& Smiljanić, K.. (2021). Enterocytes in Food Hypersensitivity Reactions. in Animals
MDPI., 11(9), 2713.
https://doi.org/10.3390/ani11092713

Krstić-Ristivojević M, Apostolović D, Smiljanić K. Enterocytes in Food Hypersensitivity Reactions. in Animals. 2021;11(9):2713.
doi:10.3390/ani11092713 .

Krstić-Ristivojević, Maja, Apostolović, Danijela, Smiljanić, Katarina, "Enterocytes in Food Hypersensitivity Reactions" in Animals, 11, no. 9 (2021):2713,
https://doi.org/10.3390/ani11092713 . .

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  - 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  - JOUR
AU  - Protić-Rosić, Isidora
AU  - Nešić, Andrijana
AU  - Lukić, Ivana
AU  - Miljković, Radmila
AU  - Popović, Dragan
AU  - Atanasković-Marković, Marina
AU  - Stojanović, Marijana
AU  - Gavrović-Jankulović, Marija
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/630
AB  - Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.
PB  - Elsevier
T2  - Molecular Immunology
T1  - Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion
EP  - 67
SP  - 58
VL  - 138
DO  - 10.1016/j.molimm.2021.06.015
ER  - 

@article{
author = "Protić-Rosić, Isidora and Nešić, Andrijana and Lukić, Ivana and Miljković, Radmila and Popović, Dragan and Atanasković-Marković, Marina and Stojanović, Marijana and Gavrović-Jankulović, Marija",
year = "2021",
abstract = "Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.",
publisher = "Elsevier",
journal = "Molecular Immunology",
title = "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion",
pages = "67-58",
volume = "138",
doi = "10.1016/j.molimm.2021.06.015"
}

Protić-Rosić, I., Nešić, A., Lukić, I., Miljković, R., Popović, D., Atanasković-Marković, M., Stojanović, M.,& Gavrović-Jankulović, M.. (2021). Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology
Elsevier., 138, 58-67.
https://doi.org/10.1016/j.molimm.2021.06.015

Protić-Rosić I, Nešić A, Lukić I, Miljković R, Popović D, Atanasković-Marković M, Stojanović M, Gavrović-Jankulović M. Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology. 2021;138:58-67.
doi:10.1016/j.molimm.2021.06.015 .

Protić-Rosić, Isidora, Nešić, Andrijana, Lukić, Ivana, Miljković, Radmila, Popović, Dragan, Atanasković-Marković, Marina, Stojanović, Marijana, Gavrović-Jankulović, Marija, "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion" in Molecular Immunology, 138 (2021):58-67,
https://doi.org/10.1016/j.molimm.2021.06.015 . .

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  - CHAP
AU  - Prodić, Ivana
AU  - Smiljanić, Katarina
AU  - Radosavljević, Jelena
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/765
AB  - Common properties of food allergens are prominent resistance to heat treatment and enzyme proteolysis. Stability of the proteins upon gastrointestinal proteolysis of food highly correlates with its allergenic potential. At this moment, the scientific community is putting a lot of effort to connect the available knowledge on the structure and function of food proteins, with stability to proteolysis in order to provide the most reliable prediction tool for allergenicity of novel proteins. Moreover, choosing the conditions under which gastrointestinal proteolysis is simulated may profoundly affect the results of assays and allergenicity assessment. At the beginning of research, for the link between allergenicity and proteolytic stability, purified allergens were used. However, this approchad was proved to be prone to production of erroneous data, since the proteolytic stability of purified proteins was frequently affected by the methodology used for protein purification and the ratio of protens to digestive enzymes used in the assays. Nowadays, the scientific community thrives to establish in vitro digestion protocols that mimic physiological conditions and take into account complex compositon of the food. New studies support this tendency, since it was shown that the presence of various biomolecules in food matrix affects the proteolysis in the simulated gastrointestinal conditions. On top of that, survival of intact proteins upon proteolysis seems not to be necessary, but frequently protein fragments of higher molecular weight with partially preserved structure might be enough to elicit allergic reaction in sensitized individuals.
PB  - New York : Nova Science Publishers
T2  - A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era
T1  - Food Allergens’ Susceptibility to Proteolysis
SP  - 220
VL  - 6
UR  - https://hdl.handle.net/21.15107/rcub_intor_765
ER  - 

@inbook{
author = "Prodić, Ivana and Smiljanić, Katarina and Radosavljević, Jelena",
year = "2020",
abstract = "Common properties of food allergens are prominent resistance to heat treatment and enzyme proteolysis. Stability of the proteins upon gastrointestinal proteolysis of food highly correlates with its allergenic potential. At this moment, the scientific community is putting a lot of effort to connect the available knowledge on the structure and function of food proteins, with stability to proteolysis in order to provide the most reliable prediction tool for allergenicity of novel proteins. Moreover, choosing the conditions under which gastrointestinal proteolysis is simulated may profoundly affect the results of assays and allergenicity assessment. At the beginning of research, for the link between allergenicity and proteolytic stability, purified allergens were used. However, this approchad was proved to be prone to production of erroneous data, since the proteolytic stability of purified proteins was frequently affected by the methodology used for protein purification and the ratio of protens to digestive enzymes used in the assays. Nowadays, the scientific community thrives to establish in vitro digestion protocols that mimic physiological conditions and take into account complex compositon of the food. New studies support this tendency, since it was shown that the presence of various biomolecules in food matrix affects the proteolysis in the simulated gastrointestinal conditions. On top of that, survival of intact proteins upon proteolysis seems not to be necessary, but frequently protein fragments of higher molecular weight with partially preserved structure might be enough to elicit allergic reaction in sensitized individuals.",
publisher = "New York : Nova Science Publishers",
journal = "A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era",
booktitle = "Food Allergens’ Susceptibility to Proteolysis",
pages = "220",
volume = "6",
url = "https://hdl.handle.net/21.15107/rcub_intor_765"
}

Prodić, I., Smiljanić, K.,& Radosavljević, J.. (2020). Food Allergens’ Susceptibility to Proteolysis. in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era
New York : Nova Science Publishers., 6, 220.
https://hdl.handle.net/21.15107/rcub_intor_765

Prodić I, Smiljanić K, Radosavljević J. Food Allergens’ Susceptibility to Proteolysis. in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era. 2020;6:220.
https://hdl.handle.net/21.15107/rcub_intor_765 .

Prodić, Ivana, Smiljanić, Katarina, Radosavljević, Jelena, "Food Allergens’ Susceptibility to Proteolysis" in A Closer Look at Proteolysis: Biochemistry and Molecular Biology in the Post Genomic Era, 6 (2020):220,
https://hdl.handle.net/21.15107/rcub_intor_765 .

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 .