Kochevar J
Cancer. 1987 Jun 15;59(12):2031-6. doi: 10.1002/1097-0142(19870615)59:12<2031::aid-cncr2820591211>3.0.co;2-0.
BALB/c mice were hyperimmunized with ACHN (ATCC CRL 1611, American Type Culture Collection, Rockville, Maryland), a stable in vitro cell line derived from a malignant pleural effusion in a 22-year-old man with renal cell carcinoma. The hyperimmune spleen cells were fused with NS-1 murine myeloma cells using polyethylene glycol. Hybridoma supernatants were screened for the presence of IgG reactive with detergent extracts of ACHN and nonreactive with detergent extracts of normal kidney tissue. A stable, rapidly growing clone named 5F4 was isolated. Supernatant from 5F4 was used as a primary antibody preparation for avidin-biotin complex immunoperoxidase staining of multiple cases of renal cell carcinoma, normal tissues, and other tumors. 5F4 produced IgG which reacted with a cytoplasmic structure in paraffin-embedded sections of all renal cell carcinomas tested. There was occasional, weak, granular, cytoplasmic staining of isolated tubular lining cells in adjacent normal kidney.
用ACHN(ATCC CRL 1611,美国典型培养物保藏中心,马里兰州罗克维尔)对BALB/c小鼠进行超免疫,ACHN是一种稳定的体外细胞系,源自一名22岁肾细胞癌男性患者的恶性胸腔积液。使用聚乙二醇将超免疫脾细胞与NS-1鼠骨髓瘤细胞融合。筛选杂交瘤上清液中是否存在与ACHN去污剂提取物反应且与正常肾组织去污剂提取物不反应的IgG。分离出一个稳定、快速生长的克隆,命名为5F4。5F4的上清液用作一抗制剂,用于对多例肾细胞癌、正常组织和其他肿瘤进行抗生物素蛋白-生物素复合物免疫过氧化物酶染色。5F4产生的IgG与所有测试的肾细胞癌石蜡包埋切片中的细胞质结构发生反应。相邻正常肾脏中偶尔有孤立的肾小管衬里细胞出现微弱的、颗粒状的细胞质染色。