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

YanMin Chen; Yin Bai; XiaoChen Guo; WenFei Wang; Qi Zheng; FuXiang Wang; Dejun Sun; DeShan Li; GuiPing Ren; JieChao Yin

doi:10.1016/j.biologicals.2016.05.003

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

Biologicals. 2016 Jul;44(4):271-275. doi: 10.1016/j.biologicals.2016.05.003. Epub 2016 May 30.

Authors

YanMin Chen¹, Yin Bai², XiaoChen Guo³, WenFei Wang⁴, Qi Zheng⁵, FuXiang Wang⁶, Dejun Sun⁷, DeShan Li⁸, GuiPing Ren⁹, JieChao Yin¹⁰

Affiliations

¹ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
² 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: [email protected].
³ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
⁴ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
⁵ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
⁶ Harbin Medical University, The Fourth Affiliated Hospital, Nangang District, Harbin 150001, People's Republic of China. Electronic address: [email protected].
⁷ Harbin Pharmaceutical Group Bio-vaccine Co., Ltd., Harbin, 150000, People's Republic of China. Electronic address: [email protected].
⁸ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
⁹ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].
¹⁰ College of Life Sciences, Northeast Agricultural University, Harbin 150030, People's Republic of China. Electronic address: [email protected].

PMID: 27255707
DOI: 10.1016/j.biologicals.2016.05.003

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.

Keywords: Affinity improvement; Bacterial display technology; CDR3 mutant library; Hepatitis B virus; Neutralizing activity; PreS1.

MeSH terms

Amino Acid Sequence
Antibodies, Neutralizing / genetics
Antibodies, Neutralizing / immunology*
Antibody Affinity / immunology
Antibody Specificity / immunology
Cell Line, Tumor
Complementarity Determining Regions / genetics
Complementarity Determining Regions / immunology
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Hep G2 Cells
Hepatitis B Antibodies / genetics
Hepatitis B Antibodies / immunology*
Hepatitis B Surface Antigens / genetics
Hepatitis B Surface Antigens / immunology*
Humans
Immunoglobulin Heavy Chains / genetics
Immunoglobulin Heavy Chains / immunology
Immunoglobulin Light Chains / genetics
Immunoglobulin Light Chains / immunology
Immunoglobulin Variable Region / genetics
Immunoglobulin Variable Region / immunology
Mutation
Peptide Library
Protein Precursors / genetics
Protein Precursors / immunology*
Single-Chain Antibodies / genetics
Single-Chain Antibodies / immunology*

Substances

Antibodies, Neutralizing
Complementarity Determining Regions
Hepatitis B Antibodies
Hepatitis B Surface Antigens
Immunoglobulin Heavy Chains
Immunoglobulin Light Chains
Immunoglobulin Variable Region
Peptide Library
Protein Precursors
Single-Chain Antibodies
presurface protein 1, hepatitis B surface antigen