摘要
采用羟基磷灰石 (HAP)作为填料 ,以pH 6 8的磷酸盐缓冲液为流动相 ,用制备型高效液相色谱来分离提纯基因重组白细胞介素 2 (IL 2 ) ,通过SDS 聚丙烯酰胺凝胶 (SDS PAGE)电泳测定IL 2的纯度 ,测定其活性为1× 1 0 6 U/mg。该法对IL 2的纯化、含量测定及活性测定等效果良好。
Interleukin 2 (IL 2)which is produced by T lymphocytes is a protein molecule with the molecular weight of 15 420 Daltons. It has good functions in curing a lot of diseases such as cancer and AIDS even with a small quantity A new method for purification of IL 2 by preparative high performance liquid chromatography is described It was the first time to use hydroxyapatite as the packing to separate IL 2 IL 2 has one disulphide bridge (between 58th Cys and 105th Cys )and one free cysteine In order to prevent aggregation and formation of wrong disulphide bond, 50 μmol/L Cu\+\{2+\} and 1 5 mol/L guanidine hydrochloride was used for autoxidation and denaturing Then we used mainly two chromatographic steps: First, Sephadex G 25 Gel permeation chromatography was used to separate guanidine hydrochloride and small molecules Second, large scale preparative HPLC was used to separate impure protein from IL 2 Gradient elution was performed with phosphate buffer (pH 6 8) Its purity was examined by SDS PAGE Its activity reached 1×10\+6 U/mg by CTLL 2 cell MTT method The results showed that it is a suitable method for the large scale purification of IL 2
出处
《色谱》
CAS
CSCD
北大核心
2000年第4期325-327,共3页
Chinese Journal of Chromatography
