Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profling Microbial Glycans
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2022
Authors
Dragačević, Luka
Lopandić, Zorana

Gavrović-Jankulović, Marija

Živković, Irena
Blagojević, Veljko
Polović, Natalija

Minić, Rajna

Article (Published version)

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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 demonstrate...d 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.
Keywords:
Glycosylation / Microorganisms / Plant lectins / Salmonella Lactobacillus / YeastsSource:
Applied Biochemistry and Biotechnology, 2022, 194, 2047-2060Publisher:
- Springer
Funding / projects:
- Ministry of Education, Science and Technological Development of the Republic of Serbia (No. 451-03-9/2021-14)
Note:
- Supplementary material: https://cherry.chem.bg.ac.rs/handle/123456789/4888
Related info:
- Referenced by
https://cherry.chem.bg.ac.rs/handle/123456789/4888
DOI: 10.1007/s12010-021-03772-w
ISSN: 0273-2289
PubMed: 35013926
WoS: 000741601700005
Scopus: 2-s2.0-85122876661
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Hemijski fakultet / Faculty of ChemistryTY - 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://cherry.chem.bg.ac.rs/handle/123456789/4887 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 T2 - Applied Biochemistry and BiotechnologyAppl Biochem Biotechnol T1 - Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profling Microbial Glycans VL - 194 SP - 2047 EP - 2060 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, Applied Biochemistry and BiotechnologyAppl Biochem Biotechnol", title = "Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profling Microbial Glycans", volume = "194", pages = "2047-2060", 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 Profling 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 Profling 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 Profling Microbial Glycans" in Applied Biochemistry and Biotechnology, 194 (2022):2047-2060, https://doi.org/10.1007/s12010-021-03772-w . .