title: [Construction, expression, and identification of the gene of human anti-prostate specific membrane antigen single-chain antibody].
abstract: OBJECTIVE
  To construct, express and purify human fusion proteins composed of a single-chain antibody fragment scFv that recognizes the prostate specific membrane antigen (PSMA) protein, Fdt, HA2 and tp, and to analyze the binding activity of the expressed fusion proteins.
  
  
  METHODS
  The fusion protein genes scFv, scFv-tp, and scFv-Fdt-HA2-tp were amplified by PCR, and the genes obtained were then cloned into the expression vector pET28 and expressed in E. coli BL21. The expressed products were identified by SDS-PAGE and Western blot and purified with Ni(2+)-NTA chelating agarose. The antigen-binding activity of the fusion proteins was determined by ELISA.
  
  
  RESULTS
  The human anti-PSMA fusion gene was successfully constructed and expressed in M15 as the inclusion body after induced with IPTG. All the target proteins expressed could bind the PSMA antigen.
  
  
  CONCLUSIONS
  Fusion proteins can specifically bind the PSMA antigen. This finding contributes to the study of the targeted delivery of siRNA.
authors.affiliation: Department of Urology, No. 323 Hospital of Chinese People's Liberation Army, Xi'an, Shaanxi, China. suys2006@163.com
authors.forename: Yan-Sheng
authors.initials: YS
authors.lastname: Su
authors.forename: Xiao-Liang
authors.initials: XL
authors.lastname: Fu
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
authors.forename: Qian-Yun
authors.initials: QY
authors.lastname: Wang
authors.forename: Na
authors.initials: N
authors.lastname: Liu
authors.forename: Hong-Bing
authors.initials: HB
authors.lastname: Jia
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Qin
authors.forename: Wei-Hong
authors.initials: WH
authors.lastname: Wen
authors.forename: He
authors.initials: H
authors.lastname: Wang
mesh_terms: D000954:Antigens, Surface / Q000276:immunology*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D043425:Glutamate Carboxypeptidase II / Q000276:immunology*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology
chemical_list: D000954:Antigens, Surface; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II