[One-step purification of monoclonal antibody against gp130 by cation exchange liquid chromatography].
OBJECTIVE
To develop a one-step purification method of anti-gp130 monoclonal antibody (mAb) B-S12 from mouse ascites.


METHODS
After filtrated by centrifugation, the ascites sample was loaded on a cation exchange column and purified by using ion-strength gradient elution buffer. The effects of pH of the loading buffer and ion strength gradients of the elution buffer on the purity of antibody obtained were investigated. The antibody's biological activity was tested by MTT colorimetry.


RESULTS
It was shown that the mAb B-S12 with a purity of over 90% could be achieved by using 20 mmol/L HEPES buffer (pH 4.0) as loading buffer and 0-1.0 mol/L NaCl as elution buffer. The total recovery rate of the mAb was 52%. The purified antibody could stimulate the proliferation of XG-2 cell line.


CONCLUSIONS
The established one-step purification method was simple and suitable for purification of mAb B-S12.
Ma
Hong-bing
HB
Institute of Medical Biotechnology, Suzhou University, Suzhou 215007, China. ma_hongbing@yahoo.com.cn
Zhuang
Yu-mei
YM
Xu
Ying
Y
Yu
Jian-feng
JF
Liu
Lin
L
Li
Wen-xiang
WX
Zhang
Xue-guang
XG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification*; D001201:Ascites / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation; D002852:Chromatography, Ion Exchange / Q000379:methods*; D050822:Cytokine Receptor gp130 / Q000276:immunology*; D051379:Mice
D000911:Antibodies, Monoclonal; D050822:Cytokine Receptor gp130