HLA—DR分子的分离纯化

目的：纯化HLA－DR。方法：纯化抗HLA－DR单克隆抗体1243并与CNBr活化的Sepharose4B偶联亲和柱，应用免疫亲和层析方法纯化HLA－DR分子。结果；从5g HLA-DR阳性的人B淋巴瘤细胞系RAJI裂解液中得到20μHLA-DR分子。它们以二聚体形式存在，能与构象依赖型单抗L243进行结合，煮沸变性后分开为α和β两条单链。这为MHCⅡ类分子抗原表位研究奠定了基础。

Purification of HLA DR molecules

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.