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Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profling Microbial Glycans

Authorized Users Only
2022
Authors
Dragačević, Luka
Lopandić, Zorana
Gavrović-Jankulović, Marija
Živković, Irena
Blagojević, Veljko
Polović, Natalija
Minić, Rajna
Article (Published version)
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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 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 / Yeasts
Source:
Applied Biochemistry and Biotechnology, 2022, n/a
Publisher:
  • 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: 1559-0291

PubMed: 35013926

WoS: 000741601700005

Scopus: 2-s2.0-85122876661
[ Google Scholar ]
URI
http://cherry.chem.bg.ac.rs/handle/123456789/4887
Collections
  • Publikacije
Institution/Community
Hemijski fakultet
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://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  - n/a
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 = "n/a",
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., n/a.
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;n/a.
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, n/a (2022),
https://doi.org/10.1007/s12010-021-03772-w . .

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