title: Selection of affinity-improved neutralizing human scFv against HBV PreS1 from CDR3 VH/VL mutant library.
abstract: 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.
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: 1320775999@qq.com.
authors.forename: YanMin
authors.initials: Y
authors.lastname: Chen
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China; The 211th Hospital of People's Liberation Army, Harbin 150080, People's Republic of China. Electronic address: 136362592@qq.com.
authors.forename: Yin
authors.initials: Y
authors.lastname: Bai
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: 676553202@qq.com.
authors.forename: XiaoChen
authors.initials: X
authors.lastname: Guo
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: 40942724@qq.com.
authors.forename: WenFei
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: 597486255@qq.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: Harbin Medical University, The Fourth Affiliated Hospital, Nangang District, Harbin 150001, People's Republic of China. Electronic address: wangfuxiang999@163.com.
authors.forename: FuXiang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Harbin Pharmaceutical Group Bio-vaccine Co., Ltd., Harbin, 150000, People's Republic of China. Electronic address: sdj711@sina.com.
authors.forename: Dejun
authors.initials: D
authors.lastname: Sun
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: deshanli@163.com.
authors.forename: DeShan
authors.initials: D
authors.lastname: Li
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: 51670277@qq.com.
authors.forename: GuiPing
authors.initials: G
authors.lastname: Ren
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: yinjiechao@126.com.
authors.forename: JieChao
authors.initials: J
authors.lastname: Yin
mesh_terms: D000595:Amino Acid Sequence; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D045744:Cell Line, Tumor; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006510:Hepatitis B Antibodies / Q000235:genetics / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D009154:Mutation; D019151:Peptide Library; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D022801:Complementarity Determining Regions; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011498:Protein Precursors; D057127:Single-Chain Antibodies; C050806:presurface protein 1, hepatitis B surface antigen
keywords: Affinity improvement; Bacterial display technology; CDR3 mutant library; Hepatitis B virus; Neutralizing activity; PreS1