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Isolation of human antibodies against hepatitis E virus from phage display library by immobilized metal affinity chromatography

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A capping-independent function of MePCE in stabilizing 7SK snRNA and facilitating the assembly of 7SK snRNP.htm (390bytes)
Date
2008
Author
Chen, Ying-Wei
Luo, Wen-Xin
Wang, Ming-Qiao
Wang, Jin
Li, Li-Feng
Yuan, Quan
Zhang, Jun
张军
Xia, Ning-Shao
夏宁邵
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  • 生命科学-已发表论文 [5901]
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Abstract
Objective To isolate human antibodies against hepatitis E virus from phage display library by a new method of panning phage antibody library based on immobilized metal affinity chromatography (IMAC). Methods Phage antibody library was allowed to mix with hex-His tagged expressed HEV specific antigen, NE2, in solution for adequate binding before affinity resin for hex-His was added. The non-specific phage antibodies were removed by extensive washing and the specific bound phage antibodies could then be eluted to infect TG1 or repeat the binding process for subsequent rounds of purification. The specificity of the selected human antibodies were tested by antigen competitive ELISA, human sera blocking ELISA, scFv expression, and sequence analysis. Results His-NE2 specific recombinant phages were successfully enriched after panning procedure. Two individual phage clones, 126 and 138, showed 50% inhibition in NE2 antigen competition ELISA and obvious blocking effect by HEV positive serum in blocking ELISA. Soluble scFv of 126, 138 bound to NE2 specifically. Conclusion Two specific human phage antibodies against hepatitis E virus (HEV) from phage display library were isolated by immobilized metal affinity chromatography. The immobilized metal affinity chromatography applied to phage antibody selection was a helpful supplement to the selection in solution.
Citation
Biomed Environ Sci. 2007 Dec;20(6):488-94.
URI
https://dspace.xmu.edu.cn/handle/2288/10480

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