论文部分内容阅读
经同源重组我们构建了同时含抗乙肝病毒表面抗原（ＨＢｓＡｇ）嵌合抗体重、轻链基因的双重组病毒ＢａｃＨＬ４．２。利用双重感染和双重组病毒感染秋粘虫Ｓｆ９细胞，ＳＤＳ－ＰＡＧＥ结果说明嵌合抗体重、轻链同时在胞内得到了表达。Ｗｅｓｔｅｒｎｂｌｏｔ分析表明两种情况均能在胞内组装成Ｈ２Ｌ２完整免疫球蛋白分子。ＥＬＩＳＡ和功能性免疫印迹检测证明重组嵌合抗体与父本鼠源单抗ＯＨ３一样能特异性识别具天然构象的ＨＢｓＡｇ，而不识别经ＳＤＳ变性的ＨＢｓＡｇ。
After homologous recombination, we constructed BacHL4.2, a dual baculovirus that contains both the heavy and light chain genes of chimeric antibody against hepatitis B virus surface antigen (HBsAg). Sf9 cells were infected with double and dual viruses. The results of SDS-PAGE showed that the chimeric antibody was heavy and the light chain was expressed in the cytoplasm at the same time. Westernblot analysis showed that both cases could be assembled intracellularly into H2L2 intact immunoglobulin molecules. ELISA and Western blotting demonstrated that recombinant chimeric antibodies, like the paternal murine mAb OH3, specifically recognized HBsAg with native conformation, but not SDS-denatured HBsAg.