Yokoyama Wayne M
University of California School of Medicine, San Francisco, California, USA.
Curr Protoc Cytom. 2006 Aug;Appendix 3:Appendix 3J. doi: 10.1002/0471142956.cya03js37.
This unit describes the production of monoclonal antibodies beginning with immunization and cell fusion and selection. Support protocols are provided for screening primary hybridoma supernatants for antibodies of desired specificity, establishment of stable hybridoma lines, cloning of these B cell lines by limiting dilution to obtain monoclonal lines, and preparation of cloning/expansion medium (thymocyte-conditioned medium).
本单元描述了单克隆抗体的生产过程,从免疫、细胞融合和筛选开始。还提供了支持方案,用于筛选初级杂交瘤上清液中具有所需特异性的抗体、建立稳定的杂交瘤细胞系、通过有限稀释法克隆这些B细胞系以获得单克隆细胞系,以及制备克隆/扩增培养基(胸腺细胞条件培养基)。