Purification of monoclonal antibody in mouse ascites by membrane affinity chromatography
10.3321/j.issn:1007-8738.2001.01.032
- VernacularTitle:用膜亲和层析法纯化小鼠腹水中的单克隆抗体
- Author:
Bing XU
;
Jiaji JIANG
- Publication Type:Journal Article
- From:
Journal of Cellular and Molecular Immunology
2001;17(1):84-86
- CountryChina
- Language:Chinese
-
Abstract:
Aim To purify monoclonal antibody in mouse ascites by a modified membrane affinity chromatography(MAC). Methods Human serum albumin HSA was adsorbed on the zeta-bind membrane (ZBM), a positively charged nylon membrane filter. Then the mouse aecites containing mAbs were filtrated through the ZBM to cause the mAbs bind to HSA adsorbed on ZBM. Finally,the purified mAbs were dissociated from the ZBM with guanidine hydrochloride solution. Results A sheet of ZBM(diameter 50mm) absorbed with HSA filtrated by 10 mL ascites could reach its maximum mAb binding capacity after two rounds of re-fitration. The dissociation of mAb from ZBM need only one round of filtration of dissociating solution. The purified mAb displayed a single band after PAGE. Sensitivity of detedcting HSA with purified mAbs was 20-fold higher than that with unpurified ascites in dot innunogold filtration assay(DIGFA). Conclusion The modified MAC with ZBM is a much easier, time-saving and effective method for affinity purification of antibodies in ascites.
