Hall B L, Murray J H, Haspel M V, Kobrin B J
Organon Teknika Biotechnology Research Institute, Rockville, Maryland 20850.
Cancer Res. 1994 Oct 1;54(19):5178-85.
The human monoclonal antibody (mAb) 123AV16-1 was generated by Epstein-Barr virus transformation of peripheral blood lymphocytes from a colorectal patient undergoing active specific immunotherapy with an autologous tumor cell-Bacille Calmette-Guérin vaccine. Direct immunohistochemical staining of tumor and normal pairs of tissues indicated that this human IgA1, lambda 2 mAb preferentially reacted with colon tumor epithelium. To generate a recombinant derivative of this Epstein-Barr virus-transformed cell line, we isolated the expressed complete heavy and light chain genes by a novel strategy and cloned them into modified pSV2-neo and pSV2-gpt expression vectors. The recombinant 123AV16-1 human mAb was expressed in both a murine myeloma and a human-murine heteromyeloma and was secreted as both monomers and dimers. The recombinant 123AV16-1 mAb expressed by both cell lines reacted with human colon tumor xenografts in a manner similar to the mAb derived from the Epstein-Barr virus-transformed cell line, indicating that the antibody specificity was not appreciably altered during the molecular rescue, cloning, or expression.
人单克隆抗体（mAb）123AV16-1是通过对一名接受自体肿瘤细胞-卡介苗主动特异性免疫治疗的结直肠癌患者的外周血淋巴细胞进行爱泼斯坦-巴尔病毒转化而产生的。对肿瘤组织和正常组织配对样本进行直接免疫组织化学染色表明，这种人IgA1、λ2单克隆抗体优先与结肠肿瘤上皮发生反应。为了产生这种爱泼斯坦-巴尔病毒转化细胞系的重组衍生物，我们通过一种新策略分离出表达的完整重链和轻链基因，并将它们克隆到修饰的pSV2-neo和pSV2-gpt表达载体中。重组123AV16-1人单克隆抗体在鼠骨髓瘤细胞和人-鼠杂种骨髓瘤细胞中均有表达，并以单体和二聚体形式分泌。两种细胞系表达的重组123AV16-1单克隆抗体与人类结肠肿瘤异种移植物的反应方式，与源自爱泼斯坦-巴尔病毒转化细胞系的单克隆抗体相似，这表明在分子拯救、克隆或表达过程中，抗体特异性没有明显改变。
