title: Construction of a human scFv antibody library with VH regions randomized and its application.
abstract: A non-immunized human single chain variable fragment (scFv) library containing 2.5 × 10(7) individual clones was constructed from antibody variable region genes of 200 non-immunized donors. ScFv gene repertories were generated by randomly combining rearranged variable regions of heavy chain (VH) and natural occurring light chain (VL) using overlapping extension PCR (OE-PCR). Five recombinant protein antigens from different species were successfully used to select specific binders. Phage ELISA showed that the recombinant phage particle could specifically bind to non-structural protein 1 of Avian influenza virus. This method can therefore efficiently generate a phage antibody library.
authors.affiliation: School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, People's Republic of China.
authors.forename: Yunchao
authors.initials: Y
authors.lastname: Liu
authors.forename: Junbiao
authors.initials: J
authors.lastname: Chang
authors.forename: Yumei
authors.initials: Y
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Wan
authors.forename: Yinbiao
authors.initials: Y
authors.lastname: Wang
authors.forename: Gaiping
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000956:Antigens, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology*; D019151:Peptide Library; D016133:Polymerase Chain Reaction / Q000379:methods; D011485:Protein Binding; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000276:immunology
chemical_list: D000956:Antigens, Viral; C085820:INS1 protein, influenza virus; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D057127:Single-Chain Antibodies; D017361:Viral Nonstructural Proteins