Construction, expression, and identification of the gene of human anti-prostate specific membrane antigen single-chain antibody.
- Author:
Yan-Sheng SU
1
;
Xiao-Liang FU
;
Dong WANG
;
Qian-Yun WANG
;
Na LIU
;
Hong-Bing JIA
;
Wei-Jun QIN
;
Wei-Hong WEN
;
He WANG
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, Surface; immunology; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Escherichia coli; genetics; immunology; Glutamate Carboxypeptidase II; immunology; Humans; Male; Polymerase Chain Reaction; RNA, Small Interfering; administration & dosage; immunology; Recombinant Fusion Proteins; genetics; immunology; Single-Chain Antibodies; genetics; immunology
- From: National Journal of Andrology 2014;20(12):1063-1067
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSThe 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.
RESULTSThe 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.
CONCLUSIONFusion proteins can specifically bind the PSMA antigen. This finding contributes to the study of the targeted delivery of siRNA.
