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Construction and characterization of porcine single-chain fragment variable antibodies that neutralize transmissible gastroenteritis virus in vitro

Author:
Zhang, Fanqing, Chen, Yuxue, Yang, Liang, Zhu, Jianguo
Source:
Archives of virology 2019 v.164 no.4 pp. 983-994
ISSN:
0304-8608
Subject:
Escherichia coli, Transmissible gastroenteritis virus, Western blotting, antibodies, antigens, clones, diarrhea, farms, fluorescent antibody technique, genes, lymphocytes, neutralization, piglets, plasmids, protein synthesis, swine diseases, therapeutics, viruses
Abstract:
Transmissible gastroenteritis virus (TGEV) infection causes severe diarrhea in piglets and imposes a significant economic burden on pig farms. Single-chain fragment variable (scFv) antibodies effectively inhibit virus infection and could be a potential therapeutic reagent for preventing disease. In this study, a recombinant scFv antibody phage display library was constructed from peripheral blood lymphocytes of piglets infected with TGEV. The library was screened with four rounds of biopanning using purified TGEV antigen, and scFv antibodies that bound to TGEV were obtained. The scFv gene was subcloned into the pET-28a(+), and the constituted plasmid was introduced into Escherichia coli BL21 (DE3) for protein expression. All three scFv clones identified had neutralizing activity against TGEV. An immunofluorescence assay and western blot analysis demonstrated that two scFv antibodies reacted with the spike protein of TGEV. These results indicate that scFv antibodies provide protection against viral infection in vitro and may be a therapeutic candidate for both prevention and treatment of TGEV infection in swine.
Agid:
6327503