摘要
【目的】制备均一性和特异性高的抗组氨酰-tRNA合成酶抗体,为研究皮肌炎和多肌炎的检测提供方法,为制备检测试剂盒提供原料。【方法】采用常规融合和间接ELISA检测法,获取杂交瘤细胞株;Protein G Sepharose 4FF和Sephadex G50结合对含有单克隆抗体的腹水进行纯化;SDS-PAGA电泳技术进行单克隆抗体的鉴定。【结果】筛选获得了2株稳定分泌阳性抗体的杂交瘤细胞株,分别命名为W001和W002;杂交瘤细胞克隆后,100%的检测孔保持了分泌抗组氨酰-tRNA合成酶抗体的能力;腹水纯化达到了较理想的效果;该2株单克隆抗体能特异性针对组氨酰-tRNA合成酶,并能跟天然的蛋白结合。【结论】所制备的均一性和特异性高的抗组氨酰-tRNA合成酶抗体,可直接用于在ELISA、免疫印迹、免疫组化学等研究方面,达到了试验的目的。
[Objective] To produce anti-histidyl-tRNA synthetase, which enjoy high degree of homogeneity and distinction,to provide the test methods for the study of the diseases of dermatomyositis and polymyositis. Besides, to produce materials for the test liquid. [Methods] With the regular combined methods available and indirect ELISA to obtain hybridoma cell lines; to purify the ascites with monoclonal antibody in the combined use of Protein G Sepharose 4FF and Sephadex G50; to identify monoclonal antibody with the electrophoresis technique. [Results] Two hybridoma cell lines, named W001 and W002, have been found in the research, which can secrete antihistidyl-tRNA synthetase monoclonal antibody stably. After the cells of hybridoma being cloned, 100% checking holes keep the capability of secreting anti-histidyl-tRNA synthetase monoclonal antibody. And the ascites of the monoclonal antibody has been purified by Protein G Sepharose 4FF and Sephadex G50, in which the purification was satisfying. In addition, the two monoclonal antibodies showed their ability to compose mold, and to integrate with natural albumen. [Conclusion] The anti-histidyl-tRNA synthetase monoclonal antibodies in this study are homogeneous and distinctive,which can be used directly in the research of ELISA,Western Blot and Immunohistochemistry etc. Obviously, the aim of this research has been achieved.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2008年第5期606-610,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省科技攻关项目(2007A020300008-6)
