title: Primary structure and functional expression of heavy- and light-chain variable region genes of a monoclonal antibody specific for human fibrin.
abstract: The immunoglobulin heavy- and light-chain variable region (VH and VK) genes were isolated from 8E5 hybridoma cells, which secreted monoclonal antibody against human fibrin by RT-PCR. An expression vector pOPE51-8E5 was constructed for the recombinant VH-VK scFv expression. The primary sequence of the variable regions was determined. Expression product was found in the periplasmic space and inclusion bodies by SDS-PAGE and immunoblotting. It was a 30 KDa single chain fragment (scFv) with the antigen-binding specificity of the parental monoclonal antibody. A light chain shuffling with an unspecific VL did not result in a loss of fibrin binding specificity.
authors.affiliation: Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Song
authors.forename: Y
authors.initials: Y
authors.lastname: Cai
authors.forename: D
authors.initials: D
authors.lastname: Song
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: H
authors.initials: H
authors.lastname: Yuan
authors.forename: L
authors.initials: L
authors.lastname: Wang
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.forename: H
authors.initials: H
authors.lastname: Lin
authors.forename: F
authors.initials: F
authors.lastname: Breitling
authors.forename: S
authors.initials: S
authors.lastname: Dübel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005337:Fibrin / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping
chemical_list: D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D005337:Fibrin