Kozarsky K F, Li L L, Schaller J, Kaminski M S, Claflin J L, Fox D A
Department of Internal Medicine, University of Michigan, Ann Arbor 48109-0531, USA.
Hybridoma. 1995 Dec;14(6):597-601. doi: 10.1089/hyb.1995.14.597.
Alteration of monoclonal antibody isotype is desirable for a variety of purposes, including obtaining an improved reagent for investigative or therapeutic use. A variety of approaches for isotype switching, particularly from IgM to various IgG subclasses, have been described. Antibodies that recognize carbohydrate determinants on glycoproteins, glycolipids, or polysaccharides are generally of the IgM isotype. This includes all available antibodies to the human CD60 antigen, a determinant with cell coactivating properties described on a subset of T lymphocytes and on other cell types. In this report a new method for monoclonal antibody isotype switching is presented. A plasmid containing the VH regions of anti-CD60 linked to C gamma 1 was transfected into a spontaneously arising variant of the CD60 hybridoma that produced kappa light chain but no heavy chain. This transfected hybridoma line maintains stable production of useful quantities of IgG1 monoclonal anti-CD60 in vitro and in vivo.
出于多种目的，包括获得用于研究或治疗的改良试剂，单克隆抗体同种型的改变是很有必要的。已经描述了多种同种型转换方法，特别是从IgM转换为各种IgG亚类。识别糖蛋白、糖脂或多糖上碳水化合物决定簇的抗体通常为IgM同种型。这包括所有针对人CD60抗原的可用抗体，CD60抗原是一种在一部分T淋巴细胞和其他细胞类型上描述的具有细胞共激活特性的决定簇。在本报告中，提出了一种单克隆抗体同种型转换的新方法。将含有与Cγ1连接的抗CD60 VH区的质粒转染到CD60杂交瘤的自发产生变体中，该变体产生κ轻链但不产生重链。这种转染的杂交瘤细胞系在体外和体内均能稳定产生大量有用的IgG1单克隆抗CD60。
