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Tytuł:
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Epitopes prediction for microcystin-LR by molecular docking.
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Autorzy:
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Liu Y; School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China. Electronic address: .
Liu S; School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Xu C; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Lin M; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Li Y; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Shen C; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Liang Y; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Sun X; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
Wang D; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China. Electronic address: .
Lü P; School of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
Liu X; School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China; Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing 210014, China.
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Źródło:
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Ecotoxicology and environmental safety [Ecotoxicol Environ Saf] 2021 Dec 20; Vol. 227, pp. 112925. Date of Electronic Publication: 2021 Oct 28.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: Amsterdam, Netherlands : Elsevier
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MeSH Terms:
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Microcystins*
Animals ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; Marine Toxins ; Mice ; Molecular Docking Simulation
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Contributed Indexing:
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Keywords: Epitope; Microcystin-LR; Molecular docking; Single chain variable fragment
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Substance Nomenclature:
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0 (Epitopes)
0 (Marine Toxins)
0 (Microcystins)
EQ8332842Y (cyanoginosin LR)
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Entry Date(s):
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Date Created: 20211030 Date Completed: 20211110 Latest Revision: 20211110
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Update Code:
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20240105
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DOI:
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10.1016/j.ecoenv.2021.112925
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PMID:
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34717216
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Microcystin-LR (MC-LR) is one of the most worldwide harmful cyanobacterial toxins. A lots of antibodies against MC-LR have been generated and characterized. However, the knowledge about the epitopes of MC-LR was still limited. The objective of this study was to analyze the epitopes of MC-LR and demonstrate the binding mode of MC-LR with its antibody. The variable genes of a mouse hybridoma cell line (Mab5H1-3B3) raised against MC-LR have been cloned and assembled in a single chain variable fragment (scFv), and then soluble expressed in E.coli BL21. Based on the scFv, the IC 50 and IC 10 for MC-LR were determined to be 7.45 nM and 0.30 nM by competitive ELISA. And the scFv also showed 115% and 112% cross-reactivities to MC-RR and MC-YR, and 59% to MC-LA. By molecular docking, the binding mode between MC-LR and its scFv was demonstrated. A hydrogen bond interaction was observed between the carbonyl group of Adda 5 residue of MC-LR and its scFv, and the guanidyl group of Arg 4 residue and phenyl group of Adda 5 residue of MC-LR were also involved in the interaction. These predicted epitopes were supported by antibody cross-reactivity data. By comparing the antibody informatics of MC-LR scFv with its predicted paratopes, VH-CDR1 was crucial for MC-LR binding, and its specificity could be tuned by engineering in Vκ-CDR1 and Vκ-CDR3. These information would be useful for the hapten design for microcystins or improving the properties of MC-LR scFv in vitro.
(Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)