title: Selection of bisphenol A - single-chain antibodies from a non-immunized mouse library by ribosome display.
abstract: 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 (KDS) of one clone was 1.76μM 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.
authors.affiliation: Tianjin Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Baoan
authors.initials: B
authors.lastname: Ning
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Jialei
authors.initials: J
authors.lastname: Bai
authors.affiliation: Tianjin Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Siming
authors.initials: S
authors.lastname: Sun
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Guimin
authors.initials: G
authors.lastname: Li
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Xianjun
authors.initials: X
authors.lastname: Fan
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Jianqing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Institute of Health and Environmental Medicine, Tianjin 300050, China. Electronic address: gaozhx@163.com.
authors.forename: Zhixian
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Tianjin Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China. Electronic address: zhangjk@tust.edu.cn.
authors.forename: Juankun
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D001559:Benzhydryl Compounds / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D002681:China; D004785:Environmental Pollutants / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D004968:Estrogens, Non-Steroidal / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D015723:Gene Library; D006241:Haptens / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D010636:Phenols / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D016415:Sequence Alignment; D057127:Single-Chain Antibodies / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013154:Spleen / Q000378:metabolism; D020349:Surface Plasmon Resonance; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D004785:Environmental Pollutants; D004968:Estrogens, Non-Steroidal; D006241:Haptens; D055767:Immobilized Proteins; D008024:Ligands; D010636:Phenols; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: Expression; Ribosome display; Sequence; Single-chain variable fragments