Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
- VernacularTitle:两步串联层析法纯化鼠抗人CD80单克隆抗体4E5
- Author:
Hong-Bing MA
;
Yu-Hua QIU
;
Ran TAO
;
Wen-Xiang LI
;
Ying XU
;
Xue-Guang ZHANG
;
- Publication Type:Journal Article
- Keywords:
Anion exchange chromatography Gel filtration chromatography Monoclonal antibody Mouse ascites Purification
- From:
China Biotechnology
2006;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.
