[Prokaryotic expression and bioactivity identification of Fab against human gamma-seminoprotein].
OBJECTIVE
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.


CONCLUSIONS
Fab against gamma -seminoprotein has been expressed successfully with biological activity, which create favourable condition for further study on targeted therapy of prostate cancer.
Sun
Ji-feng
JF
Department of Nephrology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. jifeng-sun@163.com
Yang
Jie
J
Hao
Xiao-ke
XK
Zhao
Jing
J
Wen
Wei-hong
WH
Jin
Ming
M
Wang
Cheng-ji
CJ
Yang
An-gang
AG
D000818:Animals; D006801:Humans; D015151:Immunoblotting; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000235:genetics*; D007150:Immunohistochemistry; D051379:Mice; D017430:Prostate-Specific Antigen / Q000276:immunology*
D007140:Immunoglobulin Fab Fragments; D017430:Prostate-Specific Antigen