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获取针对人类细胞表面分子的单克隆抗体经常采用完整细胞免疫小鼠的方法,由于细胞表面结构复杂,因而在制备过程中产生的单抗种类繁多,这常给筛选工作带来困难.文章报道了一种使用表达人类淋巴细胞表面抗原的昆虫细胞作为免疫原获取单抗的方法.从EB病毒转化的人B细胞株中提取RNA,逆转录合成cDNA,以其为模板用PCR扩增出CD40和B7的cDNA全长片断,以杆状病毒为载体构建成CD40和B7的重组载体,将它们分别转染sf9昆虫细胞,用细胞ELISA方法证实该细胞可在其细胞表面表达CD40和B7.用上述能表达CD40和B7的sf9细胞分
Obtaining monoclonal antibodies against human cell surface molecules often use whole-cell immunization of mice, due to the complex cell surface structure, resulting in a wide range of monoclonal antibodies in the preparation process, which often makes screening difficult. A method for obtaining a monoclonal antibody using an insect cell expressing human lymphocyte surface antigen as an immunogen RNA is extracted from a human B cell line transformed with Epstein-Barr virus and cDNA is reverse transcribed and used as a template to amplify CD40 and B7 cDNA fragment was constructed using baculovirus as a recombinant vector of CD40 and B7, which were respectively transfected into sf9 insect cells, and the cells were confirmed by cell ELISA to express CD40 and B7 on their cell surface. Sf9 cells expressing CD40 and B7