van der Velden-de Groot T A, Martin J M, Beuvery E C
Department of Inactivated Viral Vaccines, National Institute for Public Health and Environmental Protection, Bilthoven, The Netherlands.
Dev Biol Stand. 1990;71:45-54.
Monoclonal antibodies (MCA) are of great value both for in vitro and in vivo diagnostic and for therapeutic application. For in vitro application relatively small amounts of antibody are required. The therapeutic application, however, requires larger quantities. For this reason different cultivation systems have been developed in the past years. In our laboratory the continuous perfusion system in routinely applied for the cultivation of hybridoma cells. Cell and product yields obtained in this homogeneous cultivation system will be described and data of the monitoring of environmental conditions as nutrients and metabolites will be presented. To obtain information on the stability of expression a spot-ELISA assay was developed to determine the ratio of producers to non producers and the incidence of switch variants. Flow cytometry was applied for analysis of the cell cycle and to follow the course of cytoplasmic and surface IgG expression. These and other test systems provide information about the stability of production of monoclonal antibodies by hybridoma cells in continuous perfusion systems.
单克隆抗体（MCA）在体外和体内诊断以及治疗应用中都具有重要价值。对于体外应用，需要相对少量的抗体。然而，治疗应用则需要大量抗体。因此，在过去几年中开发了不同的培养系统。在我们实验室，连续灌注系统常规用于杂交瘤细胞的培养。将描述在这种均匀培养系统中获得的细胞和产物产量，并呈现营养物质和代谢产物等环境条件监测的数据。为了获得关于表达稳定性的信息，开发了一种斑点酶联免疫吸附测定法来确定产生细胞与非产生细胞的比例以及转换变体的发生率。流式细胞术用于分析细胞周期并跟踪细胞质和表面IgG表达的过程。这些以及其他测试系统提供了关于杂交瘤细胞在连续灌注系统中产生单克隆抗体稳定性的信息。
