Okabe T, Kaizu T, Fujisawa M, Watanabe J, Kojima K, Yamashita T, Takaku F
Cancer Res. 1984 Nov;44(11):5273-8.
Monoclonal antibodies to membrane antigens of human small cell carcinoma of the lung were produced by fusion of P3X63/Ag8U1 mouse myeloma cells with spleen cells from BALB/c mice immunized against the intact cells of the small cell carcinomas grown in BALB/c nude mice. The hybrids were screened for antibody production using intact cells in a solid-phase radioimmunoassay or in a membrane fluorescence with a fluorescence-activated cell sorter. Four monoclonal antibodies were chosen that demonstrated reactivities with human small cell carcinoma of the lung and not with apparently normal diploid fibroblasts or lymphoblastoid cells. The antibodies designated as TFS-1 and TFS-2 rather demonstrated "pancarcinoma" reactivity, showing binding to the other types of lung cancer (adenocarcinoma, squamous cell carcinoma, and large cell carcinoma) and carcinomas derived from other organs, such as colon, pancreas, or stomach. The monoclonal antibodies TFS-3 and TFS-4 preferentially bound to small cell carcinoma cells and neuroblastoma cells, but not to non-small cell carcinomas (adenocarcinoma, squamous cell, or large cell). Especially, TFS-4 did not bind to a variety of other normal or malignant cells. Immunoprecipitation of the antigens by monoclonal antibodies and sodium dodecyl sulfate:polyacrylamide gel electrophoresis revealed that they had different molecular weights.
通过将P3X63/Ag8U1小鼠骨髓瘤细胞与来自用在BALB/c裸鼠体内生长的小细胞癌完整细胞免疫的BALB/c小鼠的脾细胞融合,制备了针对人肺小细胞癌膜抗原的单克隆抗体。使用完整细胞通过固相放射免疫测定法或用荧光激活细胞分选仪进行膜荧光测定来筛选杂交瘤细胞的抗体产生情况。选择了四种单克隆抗体,它们与人肺小细胞癌有反应性,而与明显正常的二倍体成纤维细胞或淋巴母细胞无反应性。命名为TFS-1和TFS-2的抗体表现出“泛癌”反应性,与其他类型的肺癌(腺癌、鳞状细胞癌和大细胞癌)以及源自其他器官(如结肠、胰腺或胃)的癌有结合。单克隆抗体TFS-3和TFS-4优先结合小细胞癌细胞和神经母细胞瘤细胞,但不结合非小细胞癌(腺癌、鳞状细胞癌或大细胞癌)。特别是,TFS-4不与多种其他正常或恶性细胞结合。用单克隆抗体和十二烷基硫酸钠:聚丙烯酰胺凝胶电泳对抗原进行免疫沉淀显示它们具有不同的分子量。