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[Prokaryotic expression and bioactivity identification of Fab against human gamma-seminoprotein]
Ji-feng Sun1, Jie Yang, Xiao-ke Hao
1Department of Nephrology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. jifeng-sun@163.com
Aim:
To construct the expression vector containing cDNA encoding Fab against human gamma-seminoprotein and express it in E. coli.
Methods:
The genes encoding K chain and Fd against gamma-seminoprotein were acquired from pUC19-K and pBluescript KS( M13-)-Fd by restrictive enzyme digestion and then cloned into the expression vector pComb3 to construct recombinant expression vector pComb3-Fab. pComb3-Fab was transfected into and expressed in XLI-Blue.
Results:
Fab against r-semino-protein was expressed in. XLI-Blue. Western blot analysis and immunocytochemical staining demonstrated that ex-pressed Fab could specifically bind to gamma-seminoprotein.
Conclusion:
Fab against gamma -seminoprotein has been expressed successfully with biological activity, which create favourable condition for further study on targeted therapy of prostate cancer.
