[Preparation and identification of the monoclonal antibody against human endostatin].
OBJECTIVE
To prepare monoclonal antibody against human endostatin for the in depth study on the mechanism of anti-tumor effect of endostatin.


METHODS
Monoclonal antibody specific for endostatin was prepared using hybridoma technique and screened with ELISA. Ascites fluids were produced in BALB/c mice following in sequentical intraperitoneal injection of pristine and hybridoma cells. The mAb was purified by affinify chromatography with protein A sepharose CL-4B.


RESULTS
One hybridoma cell line 4E7 was established, which could produce mAb against human endostatin. The titers of mAb 4E7 in culture supernatant and ascites fluid were 1:128-1:516 and 1:10(-4)-1:10(-6), respectively. The mAb 4E7 belonged to IgG1 (lambda type). Western blot analysis showed that the mAb 4E7 could react to the human endostatin expressed by yeast and E.coli, but it had no cross reaction to other cytokines such as bFGF.


CONCLUSIONS
The mAb 4E7 is able to react specifically with human endostatin and can be used for further investigation.
Feng
Yu-mei
YM
Department of Genetic Engineering, Beijing Institute of Biotechnology, Beijing 100850, China. feifeicat@mailcity.com
Feng
Yi
Y
Zhang
Yan-hong
YH
Ren
Lie
L
Zeng
Zhao-chun
ZC
Ma
Qing-jun
QJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D043169:Endostatins / Q000276:immunology*; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D043169:Endostatins