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RA33/36抗原的纯化及相应抗体的检测
引用本文:闫成兰,李小峰,胡学芳,魏华,高圆,国华,吕志勤,李雪飞. RA33/36抗原的纯化及相应抗体的检测[J]. 中华风湿病学杂志, 2006, 10(5): 277-279,i0001
作者姓名:闫成兰  李小峰  胡学芳  魏华  高圆  国华  吕志勤  李雪飞
作者单位:030001,太原,山西医科大学第二医院风湿科
基金项目: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。

关 键 词:关节炎  类风湿  抗原  抗体  免疫印迹法  亲和层析  RA33/36抗原
收稿时间:2005-10-18
修稿时间:2005-10-18

The purification of RA33/36 antigen and detection of RA33/36 antibody
YAN Cheng-lan,LI Xiao-feng,HU Xue-fang,WEI Hua,GAO Yuan,GUO Hua,L Zhi-qin,LI Xue-fei. The purification of RA33/36 antigen and detection of RA33/36 antibody[J]. Chinese Journal of Rheumatology, 2006, 10(5): 277-279,i0001
Authors:YAN Cheng-lan  LI Xiao-feng  HU Xue-fang  WEI Hua  GAO Yuan  GUO Hua  L Zhi-qin  LI Xue-fei
Affiliation:Department of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan 030001, China
Abstract:
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 SDS-PAGE 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.
Keywords:Arthritis   rheumatoid  Antigen  Antibody  Immunoblotting  Affinity chromatography  RA33/36 antigen
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