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 Ni2+-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.Conclusion:Fusion proteins can specifically bind the PSMA antigen.This finding contributes to the study of the targeted delivery of siRNA.
Keywords：prostate specific membrane antigenscFvfusion proteintargeted delivery
Publication Date：2014-11-16
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)