title: [Cloning and expression of heavy- and light-chain variable region genes of monoclonal antibody specific for human fibrin].
abstract: OBJECTIVE
  To obtain the minimum molecule having activity to bind human fibrin.
  
  
  METHODS
  The immunoglobulin heavy- and light-chain variable region (VH and Vkappa) genes were isolated from 8E5 hybridoma cells, which secreted a monoclonal antibody against human fibrin, by RT-PCR. An expression vector pOPE51-8E5 was constructed for the recombinant VH-Vkappa expression. The transformed E. coli JM 109 cells were propagated and induced by IPTG.
  
  
  CONCLUSIONS
  Expression product was found in the periplasmic space and inclusion bodies by SDS-PAGE and immunobloting. It was a 30000 single chain fragment (scFv) with antigen-binding specificity. The yield of the scFv was 28.9% of the total bacterial proteins.
authors.affiliation: State Key Laboratory of Experimental Hematology, Institute of Hematology, CAMS and PUMC, Tianjin 300020.
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: L
authors.initials: L
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Yuan
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: H
authors.initials: H
authors.lastname: Lin
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D002460:Cell Line; D003001:Cloning, Molecular; D005337:Fibrin / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D005337:Fibrin