title: Affinity maturation of single-chain variable fragment specific for aflatoxin B(1) using yeast surface display.
abstract: As aflatoxin B1 is one of the most toxic mycotoxins, it is important to detect and to quantify aflatoxin B1 accurately by immunological methods. To enhance aflatoxin B1-binding affinity of the single-chain variable fragment, yeast surface display technique combined with fluorescence-activated cell sorting was applied. A randomly mutated scFv library was subjected to 4 rounds of fluorescence-activated cell sorting, resulting in isolation of 5 scFv variants showing an affinity improvement compared to the parental wild type scFv. The best scFv with a 9-fold improvement in affinity for aflatoxin B1 exhibited similar specificity to the monoclonal antibody. Most of the mutations in scFv-M37 were located outside of the canonical antigen-contact loops, suggesting that its affinity improvement might be driven by an allosteric effect inducing scFv-M37 to form a more favorable binding pocket for aflatoxin B1 than the wild type scFv.
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Won-Ki
authors.initials: WK
authors.lastname: Min
authors.affiliation: Department of Biomedical Science, Youngdong University, Chungbuk 370-701, Republic of Korea.
authors.forename: Sung-Gun
authors.initials: SG
authors.lastname: Kim
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 151-742, Republic of Korea. Electronic address: jhseo94@snu.ac.kr.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
mesh_terms: D016604:Aflatoxin B1 / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D001230:Aspergillus / Q000737:chemistry* / Q000254:growth & development; D005434:Flow Cytometry / Q000379:methods*; D057127:Single-Chain Antibodies / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D057127:Single-Chain Antibodies; D016604:Aflatoxin B1
keywords: Aflatoxin B(1); Fluorescence-activated cell sorting; Single-chain variable fragment; Yeast surface display