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

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• 作者：YanMin Chen^a ; ^{1320775999@qq.com" class="auth_mail" title="E-mail the corresponding author} ; Yin Bai^a ; ^b ; ^{136362592@qq.com" class="auth_mail" title="E-mail the corresponding author} ; XiaoChen Guo^a ; ^{676553202@qq.com" class="auth_mail" title="E-mail the corresponding author} ; WenFei Wang^a ; ^{40942724@qq.com" class="auth_mail" title="E-mail the corresponding author} ; Qi Zheng^a ; ^{597486255@qq.com" class="auth_mail" title="E-mail the corresponding author} ; FuXiang Wang^c ; ^{wangfuxiang999@163.com" class="auth_mail" title="E-mail the corresponding author} ; Dejun Sun^d ; ^{sdj711@sina.com" class="auth_mail" title="E-mail the corresponding author} ; DeShan Li^a ; ^{deshanli@163.com" class="auth_mail" title="E-mail the corresponding author} ; GuiPing Ren^a ; ^{51670277@qq.com" class="auth_mail" title="E-mail the corresponding author} ; JieChao Yin^a ; ^{yinjiechao@126.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Hepatitis B virus ; PreS1 ; CDR3 mutant library ; Affinity improvement ; Bacterial display technology ; Neutralizing activity
• 刊名：Biologicals
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：44
• 期：4
• 页码：271-275
• 全文大小：1041 K

文摘

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.