Yokoyama Wayne M
Howard Hughes Medical Institute and Washington University School of Medicine, St. Louis, Missouri, USA.
Curr Protoc Mol Biol. 2008 Jul;Chapter 11:Unit 11.10. doi: 10.1002/0471142727.mb1110s83.
This unit details methods for production of monoclonal antibodies. Two methods are given for production of hybridoma supernatants, including one for large-scale production. A protocol for large-scale production of hybridomas or cell lines is presented for use in isolation of cellular proteins. Finally, a method is given for producing and obtaining mouse ascites fluid containing monoclonal antibody. Recommendations developed by the Committee on Methods of Producing Monoclonal Antibodies (Institute of Laboratory Animal Research, National Research Council) on the use of animals for producing monoclonal antibodies are also discussed. These recommendations are designed to foster the judicious use of animals and to strongly encourage the use of in vitro methods whenever possible.
本单元详细介绍了单克隆抗体的生产方法。给出了两种制备杂交瘤细胞上清液的方法,包括一种大规模生产的方法。还介绍了一种用于大规模生产杂交瘤或细胞系以分离细胞蛋白的方案。最后,给出了一种制备和获取含单克隆抗体的小鼠腹水的方法。还讨论了生产单克隆抗体动物使用委员会(国家研究委员会实验动物研究所在)制定的关于使用动物生产单克隆抗体的建议。这些建议旨在促进对动物的合理使用,并强烈鼓励尽可能使用体外方法。