TY  - JOUR
AU  - Dragačević, Luka
AU  - Lopandić, Zorana
AU  - Gavrović-Jankulović, Marija
AU  - Živković, Irena
AU  - Blagojević, Veljko
AU  - Polović, Natalija
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/621
AB  - The surface of microorganisms is covered with carbohydrates, which makes them unique, self-sustaining glycan probes. Lectins are able to bind to these probes, and this interaction can be exploited for selecting microorganisms or novel lectins. To examine lectin-microorganism interactions, we have previously developed an enzyme-linked lectin sorbent assay (ELLSA) with whole bacterial cells. To further test the validity of this methodology, here we compare it with flow cytometry. For this purpose, we used biotinylated recombinantly produced lectin from Musa acuminata (BanLec), this lectin’s recombinantly produced chimera with green fluorescent protein (BanLec-eGFP) and a lectin from Ricinus communis (RCA120), both biotinylated and FITC labeled. Parallel testing showed equivalent results for the two methods, in terms of the presence or absence of binding, with signal intensity yielding high Pearson correlation coefficient of 0.8 for BanLec and 0.95 for RCA120. The ELLSA method demonstrated multiple advantages, such as reliability and convenience for high-throughput analysis; it also required less lectin and yielded more consistent results. As such, ELLSA proved to be a useful tool for profiling microbial glycan structures or testing novel lectins.
PB  - Springer
T2  - Applied Biochemistry and Biotechnology
T1  - Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans
EP  - 2060
SP  - 2047
VL  - 194
DO  - 10.1007/s12010-021-03772-w
ER  - 

@article{
author = "Dragačević, Luka and Lopandić, Zorana and Gavrović-Jankulović, Marija and Živković, Irena and Blagojević, Veljko and Polović, Natalija and Minić, Rajna",
year = "2022",
abstract = "The surface of microorganisms is covered with carbohydrates, which makes them unique, self-sustaining glycan probes. Lectins are able to bind to these probes, and this interaction can be exploited for selecting microorganisms or novel lectins. To examine lectin-microorganism interactions, we have previously developed an enzyme-linked lectin sorbent assay (ELLSA) with whole bacterial cells. To further test the validity of this methodology, here we compare it with flow cytometry. For this purpose, we used biotinylated recombinantly produced lectin from Musa acuminata (BanLec), this lectin’s recombinantly produced chimera with green fluorescent protein (BanLec-eGFP) and a lectin from Ricinus communis (RCA120), both biotinylated and FITC labeled. Parallel testing showed equivalent results for the two methods, in terms of the presence or absence of binding, with signal intensity yielding high Pearson correlation coefficient of 0.8 for BanLec and 0.95 for RCA120. The ELLSA method demonstrated multiple advantages, such as reliability and convenience for high-throughput analysis; it also required less lectin and yielded more consistent results. As such, ELLSA proved to be a useful tool for profiling microbial glycan structures or testing novel lectins.",
publisher = "Springer",
journal = "Applied Biochemistry and Biotechnology",
title = "Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans",
pages = "2060-2047",
volume = "194",
doi = "10.1007/s12010-021-03772-w"
}

Dragačević, L., Lopandić, Z., Gavrović-Jankulović, M., Živković, I., Blagojević, V., Polović, N.,& Minić, R.. (2022). Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. in Applied Biochemistry and Biotechnology
Springer., 194, 2047-2060.
https://doi.org/10.1007/s12010-021-03772-w

Dragačević L, Lopandić Z, Gavrović-Jankulović M, Živković I, Blagojević V, Polović N, Minić R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. in Applied Biochemistry and Biotechnology. 2022;194:2047-2060.
doi:10.1007/s12010-021-03772-w .

Dragačević, Luka, Lopandić, Zorana, Gavrović-Jankulović, Marija, Živković, Irena, Blagojević, Veljko, Polović, Natalija, Minić, Rajna, "Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans" in Applied Biochemistry and Biotechnology, 194 (2022):2047-2060,
https://doi.org/10.1007/s12010-021-03772-w . .

TY  - JOUR
AU  - Dragačević, Luka
AU  - Đorđević, Brižita
AU  - Gavrović-Jankulović, Marija
AU  - Ilić, Vesna
AU  - Kanazir, Danijela
AU  - Minić, Rajna
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/563
AB  - The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.
PB  - Springer, Dordrecht
T2  - Glycoconjugate Journal
T1  - ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin
EP  - 105
IS  - 1
SP  - 95
VL  - 37
DO  - 10.1007/s10719-019-09898-8
ER  - 

@article{
author = "Dragačević, Luka and Đorđević, Brižita and Gavrović-Jankulović, Marija and Ilić, Vesna and Kanazir, Danijela and Minić, Rajna",
year = "2020",
abstract = "The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.",
publisher = "Springer, Dordrecht",
journal = "Glycoconjugate Journal",
title = "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin",
pages = "105-95",
number = "1",
volume = "37",
doi = "10.1007/s10719-019-09898-8"
}

Dragačević, L., Đorđević, B., Gavrović-Jankulović, M., Ilić, V., Kanazir, D.,& Minić, R.. (2020). ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal
Springer, Dordrecht., 37(1), 95-105.
https://doi.org/10.1007/s10719-019-09898-8

Dragačević L, Đorđević B, Gavrović-Jankulović M, Ilić V, Kanazir D, Minić R. ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal. 2020;37(1):95-105.
doi:10.1007/s10719-019-09898-8 .

Dragačević, Luka, Đorđević, Brižita, Gavrović-Jankulović, Marija, Ilić, Vesna, Kanazir, Danijela, Minić, Rajna, "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin" in Glycoconjugate Journal, 37, no. 1 (2020):95-105,
https://doi.org/10.1007/s10719-019-09898-8 . .

TY  - CONF
AU  - Dragačević, Luka
AU  - Minić, Rajna
AU  - Michalickova, Danica
AU  - Ilić, Vesna
AU  - Đorđević, Brižita
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/568
PB  - Lippincott Williams & Wilkins, Philadelphia
C3  - Journal of Clinical Gastroenterology
T1  - Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults
EP  - S18
SP  - S18
VL  - 54
UR  - https://hdl.handle.net/21.15107/rcub_intor_568
ER  - 

@conference{
author = "Dragačević, Luka and Minić, Rajna and Michalickova, Danica and Ilić, Vesna and Đorđević, Brižita",
year = "2020",
publisher = "Lippincott Williams & Wilkins, Philadelphia",
journal = "Journal of Clinical Gastroenterology",
title = "Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults",
pages = "S18-S18",
volume = "54",
url = "https://hdl.handle.net/21.15107/rcub_intor_568"
}

Dragačević, L., Minić, R., Michalickova, D., Ilić, V.,& Đorđević, B.. (2020). Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults. in Journal of Clinical Gastroenterology
Lippincott Williams & Wilkins, Philadelphia., 54, S18-S18.
https://hdl.handle.net/21.15107/rcub_intor_568

Dragačević L, Minić R, Michalickova D, Ilić V, Đorđević B. Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults. in Journal of Clinical Gastroenterology. 2020;54:S18-S18.
https://hdl.handle.net/21.15107/rcub_intor_568 .

Dragačević, Luka, Minić, Rajna, Michalickova, Danica, Ilić, Vesna, Đorđević, Brižita, "Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults" in Journal of Clinical Gastroenterology, 54 (2020):S18-S18,
https://hdl.handle.net/21.15107/rcub_intor_568 .

TY  - CONF
AU  - Dragačević, Luka
AU  - Đorđević, Brižita
AU  - Gavrović-Jankulović, Marija
AU  - Ilić, Vesna
AU  - Minić, Rajna
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/567
PB  - Lippincott Williams & Wilkins, Philadelphia
C3  - Journal of Clinical Gastroenterology
T1  - Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa
EP  - S13
SP  - S13
VL  - 54
UR  - https://hdl.handle.net/21.15107/rcub_intor_567
ER  - 

@conference{
author = "Dragačević, Luka and Đorđević, Brižita and Gavrović-Jankulović, Marija and Ilić, Vesna and Minić, Rajna",
year = "2020",
publisher = "Lippincott Williams & Wilkins, Philadelphia",
journal = "Journal of Clinical Gastroenterology",
title = "Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa",
pages = "S13-S13",
volume = "54",
url = "https://hdl.handle.net/21.15107/rcub_intor_567"
}

Dragačević, L., Đorđević, B., Gavrović-Jankulović, M., Ilić, V.,& Minić, R.. (2020). Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa. in Journal of Clinical Gastroenterology
Lippincott Williams & Wilkins, Philadelphia., 54, S13-S13.
https://hdl.handle.net/21.15107/rcub_intor_567

Dragačević L, Đorđević B, Gavrović-Jankulović M, Ilić V, Minić R. Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa. in Journal of Clinical Gastroenterology. 2020;54:S13-S13.
https://hdl.handle.net/21.15107/rcub_intor_567 .

Dragačević, Luka, Đorđević, Brižita, Gavrović-Jankulović, Marija, Ilić, Vesna, Minić, Rajna, "Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa" in Journal of Clinical Gastroenterology, 54 (2020):S13-S13,
https://hdl.handle.net/21.15107/rcub_intor_567 .

TY  - JOUR
AU  - Pavlović, Bojana
AU  - Cvijetić, Nataša
AU  - Dragačević, Luka
AU  - Ivković, Branka
AU  - Vujić, Zorica
AU  - Kuntić, Vesna
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/469
AB  - One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.
PB  - Oxford Univ Press Inc, Cary
T2  - Journal of AOAC International
T1  - Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine
EP  - 400
IS  - 2
SP  - 396
VL  - 99
DO  - 10.5740/jaoacint.15-0201
ER  - 

@article{
author = "Pavlović, Bojana and Cvijetić, Nataša and Dragačević, Luka and Ivković, Branka and Vujić, Zorica and Kuntić, Vesna",
year = "2016",
abstract = "One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.",
publisher = "Oxford Univ Press Inc, Cary",
journal = "Journal of AOAC International",
title = "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine",
pages = "400-396",
number = "2",
volume = "99",
doi = "10.5740/jaoacint.15-0201"
}

Pavlović, B., Cvijetić, N., Dragačević, L., Ivković, B., Vujić, Z.,& Kuntić, V.. (2016). Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International
Oxford Univ Press Inc, Cary., 99(2), 396-400.
https://doi.org/10.5740/jaoacint.15-0201

Pavlović B, Cvijetić N, Dragačević L, Ivković B, Vujić Z, Kuntić V. Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International. 2016;99(2):396-400.
doi:10.5740/jaoacint.15-0201 .

Pavlović, Bojana, Cvijetić, Nataša, Dragačević, Luka, Ivković, Branka, Vujić, Zorica, Kuntić, Vesna, "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine" in Journal of AOAC International, 99, no. 2 (2016):396-400,
https://doi.org/10.5740/jaoacint.15-0201 . .