Selection of bisphenol A - single-chain antibodies from a non-immunized mouse library by ribosome display

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• 作者：Li Zhao^a ; ¹ ; Baoan Ning^b ; ¹ ; Jialei Bai^b ; Xiang Chen^a ; Yuan Peng^b ; Siming Sun^b ; Guimin Li^b ; Xianjun Fan^b ; Yuanyuan Liu^b ; Jianqing Liu^b ; Yanan Sun^b ; Zhixian Gao^b ; ^{gaozhx@163.com" class="auth_mail" title="E-mail the corresponding author} ; Juankun Zhang^a ; ^{zhangjk@tust.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Ribosome display ; Single-chain variable fragments ; Sequence ; Expression
• 刊名：Analytical Biochemistry
• 出版年：2015
• 出版时间：1 November 2015
• 年：2015
• 卷：488
• 期：Complete
• 页码：59-64
• 全文大小：1744 K

文摘

Developing reagents with high affinity and specificity are critical to detect the environmental hormones or toxicants. Ribosome display technology has been widely used in functional protein or peptide screening and in directed evolution of protein molecules in vitro. In this study, single-chain variable fragments (scFvs) against bisphenol A (BPA) were selected from a library constructed from splenocytes of non-immunized mice. After five rounds of selection, the selected scFvs bound to BPA with high affinity. Indirect competitive enzyme-linked immunosorbent assay (ELISA) was introduced to screen the antibody affinity and specificity to BPA. The equilibrium dissociation constants (KD_S) of one clone was 1.76 渭M as determined by surface plasmon resonance (SPR). This study indicated that ribosome display can isolate binders to small molecules from a non-immunized naive library without any in vivo steps and can generate recombinant antibodies efficiently and rapidly. In addition, this study provides a methodological framework for detection of small molecules using recombinant antibodies.