HLA-DR分子的分离纯化

Purification of HLA DR molecules

目的 :纯化HLA DR分子。方法 :纯化抗HLA DR单克隆抗体L2 4 3并与CNBr活化的Sepharose 4B偶联制备亲和柱 ,应用免疫亲和层析方法纯化HLA DR分子。结果 :从 5gHLA DR阳性的人B淋巴瘤细胞系RAJI裂解液中得到 2 0 μgHLA DR分子。它们以二聚体形式存在 ,能与构象依赖型单抗L2 4 3进行结合 ,煮沸变性后分开为α和 β两条单链。

Objective:To purify HLA DR molecules. Methods: Anti HLA DR antibody L243 was purified and coupled with CNBr activated Sepharose 4B gel. Immunoaffinity column was used to purify HLA DR molecules. Results:Twenty micrograms of HLA DR molecules were isolated from about 5 g Epstein Barr virus transformed human B lymphoblastoid cell line RAJI lysates by affinity chromatography. The purified HLA DR molecules existed in α/β heterodimers form and could bind to conformation dependent antibody L243. These HLA DR molecules were separated into two strands,α and β,by boiling denaturation. These results are the basis for studying MHC Ⅱ binding peptide motif and CD4 + T cell epitopes of antigens in future. [