从RPMI8866细胞制备CD23分子最佳条件及提取方法的研究

PREPARATION OF SOLUBLE CD23

建立从高表达CD23分子的RPMI8866细胞株分离制备CD23分子的两种方法:自然裂解法和木瓜蛋白酶水解法。比较了此两法的优劣,探讨了CD23回收量的影响因素,经Western blot-ting技术鉴定出制备品中18、37、42、45和86KDa的CD23片段。结果表明,上述方法简便、可靠、易行,且回收量较高。可供研究及分离其它膜分子参考。

RPMI 8866 cell line, expressing highly CD23 molecules, was used to produce soluble CD23 (sCD23).Two methods, Enzyme-digesting and Natural-splitting, were successfully established to prepare sCD23from the cell line. Optimum isolating conditions were tested and determined. Furthermore, the resultsshowed with Western blotting the five bands: 86KDa, 45KDa, 42KDa, 37KDa and 18KDa separated bySDS-PAGE cou1d specifically combine with human myloma IgE.