Lindgren J, Blaszczyk M, Atkinson B, Steplewski Z, Koprowski H
Cancer Immunol Immunother. 1986;22(1):1-7. doi: 10.1007/BF00205709.
Over 600 hybridomas were derived from the immunization of mice with live cells and aqueous extracts of the human prostatic carcinoma cell line PC3. A total of 26 hybridomas with restricted reactivities were selected, subcloned and antibodies tested on a variety of tumor and normal cells. Seven monoclonal antibodies showed reactivity for prostate cancer and other tumor cell lines, including breast carcinomas. Three of the antibodies obtained after immunization with live cells reacted with live cells only and three of the four antibodies obtained after immunization with cell extract reacted with cell extracts and spent culture media. The fourth antibody in the latter group was reactive only in the immunoperoxidase staining assay. Antibody PrS5 recognized a 90,000 molecular weight molecule from 125I-surface-labeled cells in immunoprecipitation analysis. Antibodies PrE3 and PrD8 detected a nonacid glycolipid pentasaccharide from PC3 cells and meconium, and a glycoprotein of 115,000 molecular weight from 125I-surface-labeled red blood cells. The similar patterns of reactivity in RIAs and antigen analysis suggest that antibodies PrE3 and PrD8 recognize the same molecule. The results emphasize the usefulness of immunohistochemistry in the testing of monoclonal antibodies and the impact of the form in which the antigen is presented on the resultant antibody specificity.
用人类前列腺癌细胞系PC3的活细胞和水提取物免疫小鼠,获得了600多个杂交瘤。总共筛选出26个反应性受限的杂交瘤,进行亚克隆,并在多种肿瘤细胞和正常细胞上测试抗体。七种单克隆抗体对前列腺癌和其他肿瘤细胞系有反应,包括乳腺癌。用活细胞免疫后获得的三种抗体仅与活细胞反应,用细胞提取物免疫后获得的四种抗体中的三种与细胞提取物和用过的培养基反应。后一组中的第四种抗体仅在免疫过氧化物酶染色试验中有反应。在免疫沉淀分析中,抗体PrS5从125I表面标记的细胞中识别出一种分子量为90,000的分子。抗体PrE3和PrD8从PC3细胞和胎粪中检测到一种非酸性糖脂五糖,从125I表面标记的红细胞中检测到一种分子量为115,000的糖蛋白。放射免疫分析和抗原分析中相似的反应模式表明,抗体PrE3和PrD8识别同一分子。结果强调了免疫组织化学在单克隆抗体检测中的有用性,以及抗原呈现形式对所得抗体特异性的影响。