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Selection of affinity-improved neutralizing human scFv against HBV PreS1 from CDR3 VH/VL mutant library

Author:
Chen, YanMin, Bai, Yin, Guo, XiaoChen, Wang, WenFei, Zheng, Qi, Wang, FuXiang, Sun, Dejun, Li, DeShan, Ren, GuiPing, Yin, JieChao
Source:
Biologicals 2016 v.44 no.4 pp. 271-275
ISSN:
1045-1056
Subject:
DNA primers, Escherichia coli, Homo sapiens, antibodies, binding capacity, clones, enzyme-linked immunosorbent assay, flow cytometry, globulins, hepatitis B, hepatocytes, humans, mutants, neutralization, neutralization tests, polymerase chain reaction, rapid methods, screening
Abstract:
A CDR3 mutant library was constructed from a previously isolated anti-HBV neutralizing Homo sapiens scFv-31 template by random mutant primers PCR. Then the library was displayed on the inner membrane surface in Escherichia coli periplasmic space. Seven scFv clones were isolated from the mutant library through three rounds of screening by flow cytometry. Competition ELISA assay indicates that isolated scFv fragments show more efficient binding ability to HBV PreS1 compared with parental scFv-31. HBV neutralization assay indicated that two clones (scFv-3 and 59) show higher neutralizing activity by blocking the HBV infection to Chang liver cells. Our method provides a new strategy for rapid screening of mutant antibody library for affinity-enhanced scFv clones and the neutralizing scFvs obtained from this study provide a potential alternative of Hepatitis B immune globulin.
Agid:
5537839