Mukerjee S, Nasoff M, McKnight M, Glassy M
Novopharm Biotech, Inc., San Diego, CA 92126, USA.
Hybridoma. 1998 Apr;17(2):133-42. doi: 10.1089/hyb.1998.17.133.
A human IgG1.k monoclonal antibody (MAb) designated GMA1 was developed by fusing pooled lymph node lymphocytes from cancer patients with the human lymphoblastoid cell line, SHFP-1. The GMA1 MAb reacted with several melanoma and neuroblastoma cell lines. Normal tissue derived from human brain and tumor-cell lines derived from colon, ovary, and breast were not reactive. FACS analysis performed using live cells demonstrated that the antibody recognizes a cell-surface antigen. Enzyme immunoassay (EIA) and thin layer chromatography (TLC) immunostaining with purified gangliosides indicated that the antibody has specificity for the major tumor associated gangliosides GD3, GM3, and GD2. GMA1 heavy and light chain genes were isolated by RT-PCR and a recombinant derivative of this human antibody was expressed in Chinese hamster ovary (CHO) cells. High-level antibody synthesis and secretion was achieved using a vector designed to maximize expression. FACS analysis and TLC immunostaining indicated recombinant GMA1 reacted with human tumor cell lines and gangliosides GD3, GM3, GD2 in a manner similar to the antibody produced by the hybridoma cell line, demonstrating that the specificity of the antibody was not altered during molecular cloning.
一种名为GMA1的人IgG1.k单克隆抗体（MAb）是通过将癌症患者的汇集淋巴结淋巴细胞与人淋巴母细胞系SHFP-1融合而研制的。GMA1单克隆抗体与几种黑色素瘤和神经母细胞瘤细胞系发生反应。源自人脑的正常组织以及源自结肠、卵巢和乳腺的肿瘤细胞系均无反应性。使用活细胞进行的流式细胞术分析表明，该抗体识别一种细胞表面抗原。用纯化的神经节苷脂进行酶免疫测定（EIA）和薄层色谱（TLC）免疫染色表明，该抗体对主要的肿瘤相关神经节苷脂GD3、GM3和GD2具有特异性。通过RT-PCR分离出GMA1重链和轻链基因，并在中华仓鼠卵巢（CHO）细胞中表达了这种人抗体的重组衍生物。使用旨在最大化表达的载体实现了高水平的抗体合成和分泌。流式细胞术分析和TLC免疫染色表明，重组GMA1与人肿瘤细胞系以及神经节苷脂GD3、GM3、GD2发生反应，其方式与杂交瘤细胞系产生的抗体相似，这表明该抗体的特异性在分子克隆过程中未发生改变。
