论文部分内容阅读
建立了一种同时快速分离纯化C1q及酶原形式C1r和C1s的方法。用IgG Sepharose 4B亲和层析将C1q与C1r2 s2 分离 ,接着用DEAE Sephacel离子交换层析将C1r和C1s分离 ,用C1qMcAb Sepharose 4B亲和层析将C1q进一步纯化。在分离Clr和C1s的整个过程中加入蛋白水解抑制剂PMSF和NPGB ,并控制温度在 4℃、pH 6 1、无二价阳离子的条件下 ,得到高度纯化的C1q、C1r和C1s。
A method for simultaneous rapid separation and purification of Clq and zymogen forms C1r and C1s was established. C1q was separated from C1r2s2 by affinity chromatography with IgG Sepharose 4B followed by separation of C1r and C1s by DEAE Sepharose ion exchange chromatography and C1q was further purified by C1qMcAb Sepharose 4B affinity chromatography. Highly purified C1q, C1r and C1s were obtained by adding the proteolytic inhibitors PMSF and NPGB throughout Clr and C1s isolation and controlling the temperature at 4 ° C, pH 6 1 without divalent cations.