Weidle U H, Borgya A, Mattes R, Lenz H, Buckel P
Gene. 1987;51(1):21-9. doi: 10.1016/0378-1119(87)90470-7.
We report here for the first time reconstitution and secretion of functionally active antibody in non-lymphoid cells. Expression vectors for the light and the heavy chain of a monoclonal antibody directed against creatine kinase (EC 2.7.3.2) were introduced into COS and CHO Chinese hamster ovary dhfr- cells. Introduction of the expression vectors separately gave rise to immuno-reactive material in the culture supernatants, but only cotransfection of the expression plasmids resulted in secretion of protein with immuno-reactivity against antibodies directed against mouse heavy and light chains as well as specific antigen-binding affinity, as determined by enzyme-linked immunosorbent assay. Secreted kappa and gamma chains from reconstituted antibody were characterized by immunoadsorption and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In COS cells, reconstituted antibody was transiently secreted; cotransfection of kappa and gamma chain expression plasmids with a dihydrofolate reductase (DHFR)-expression plasmid into CHO dhfr- cells gave rise to stable transformants secreting functionally active antibody.
我们首次在此报告非淋巴细胞中功能性活性抗体的重组与分泌。将针对肌酸激酶（EC 2.7.3.2）的单克隆抗体轻链和重链的表达载体导入COS细胞和中国仓鼠卵巢dhfr-细胞。单独导入表达载体可在培养上清液中产生免疫反应性物质，但只有共转染表达质粒才会导致分泌出具有针对小鼠重链和轻链抗体的免疫反应性以及通过酶联免疫吸附测定确定的特异性抗原结合亲和力的蛋白质。通过免疫吸附和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对重组抗体分泌的κ链和γ链进行了表征。在COS细胞中，重组抗体是瞬时分泌的；将κ链和γ链表达质粒与二氢叶酸还原酶（DHFR）表达质粒共转染到CHO dhfr-细胞中可产生分泌功能性活性抗体的稳定转化体。
