RA33/36抗原的纯化及相应抗体的检测  

The purification of RA33/36 antigen and detection of RA33/36 antibody

作　　者：闫成兰[1] 李小峰[1] 胡学芳[1] 魏华[1] 高圆[1] 国华[1] 吕志勤[1] 李雪飞[1] 

机构地区：[1]山西医科大学第二医院风湿科,太原030001

出　　处：《中华风湿病学杂志》2006年第5期277-279,i0001,共4页Chinese Journal of Rheumatology

基　　金：山西省自然科学基金资助项目(19991084)

摘　　要：目的纯化RA33/36抗原,观察其检测效率。方法从小鼠Ehrlich腹水癌细胞中提取RA33/36粗抗原,使用肝素-琼脂糖凝胶CL-6B对RA33/36粗抗原进行亲和层析,SDS-聚丙烯酰胺凝胶(SDS-PAGE)和免疫印迹法(IBT)检测富含RA33/36的组分,比较粗抗原和纯化抗原的检测效果。结果SDS-PAGE和IBT显示RA33/36抗原主要存在于缓冲液C的洗脱峰中,纯化抗原较粗抗原杂蛋白条带明显减少,且更加清晰,在检测RA33/36抗体时得到了相同的阳性和阴性结果。结论纯化的RA33/36抗原较粗抗原提高了抗体检测效率,可广泛应用于临床。Objective To purify the crude RA33/36 antigen and study the detecting efficiency. Methods The RA33/36 antigen were extracted from Ehrlich ascites carcinoma cells and were purified by affinity chromatography on heparin-sepharose CL-6B. Fractions containing RA33/36 were tested by SDSPAGE and IBT. The detecting efficiency between the crude RA33/36 antigen and purified antigen was compared. Results The SDS-PAGE showed that RA33/36 antibody presented mainly in the eluating fractions of buffer C. The immunoblotting showed, compared with the crude antigen, the interference bands of the purified antigen were much significantly decreased and the bands of RA33/36 antibody were more clear, but the positive rates had no difference between them. Conclusion The detecting efficiency of RA33/36 antibody is improved when the purified antigen is used for immunoblotting.

关 键 词：类风湿关节炎 抗原 抗体 免疫印迹法 亲和层析 RA33/36抗原 

分 类 号：R593.22[医药卫生—内科学]