Avner B P, Liao S K, Avner B, DeCell K, Oldham R K
Biotherapeutics Inc., Franklin, TN 37064.
J Biol Response Mod. 1989 Feb;8(1):25-36.
The purpose of this work was to create antibodies that are highly specific to epitopes on the surface of patient tumor cells that, when added together as a "cocktail," bind to greater than 99% of the patient's tumor cells. We describe the rationale and the methods used to develop new murine hybridomas that secrete monoclonal antibodies reactive with surface markers expressed on breast, lung, colon, kidney, islet cell, and miscellaneous carcinomas and melanoma. A rapid immunofluorescence method (cell concentration fluorescence immunoassay) is also described that has been developed to rapidly screen culture supernatants on viable patient tumor cells, tumor cell lines, or peripheral blood cells. We report the development of one breast, five colon, and three melanoma, three nephroma, and two pancreatic islet cell carcinoma antibodies. The breast antibody binds to 75% of the breast tumors tested thus far and to the same percentage of colon tumors as do the five colon antibodies, 77-85%. The melanoma antibodies described react with 90-100% of the melanoma and prostate cancers tested. The total process of creating the hybridomas and screening the antibodies for potential clinical usefulness has taken from 6 to 9 months to complete, including testing normal tissue reactivity by immunohistochemistry and production of gram quantities of monoclonal antibodies from ascites.
这项工作的目的是制备对患者肿瘤细胞表面表位具有高度特异性的抗体，当将这些抗体作为“鸡尾酒”组合在一起时，能与超过99%的患者肿瘤细胞结合。我们描述了开发新的鼠杂交瘤的基本原理和方法，这些杂交瘤分泌的单克隆抗体可与乳腺癌、肺癌、结肠癌、肾癌、胰岛细胞癌、其他各类癌症及黑色素瘤中表达的表面标志物发生反应。还介绍了一种快速免疫荧光方法（细胞浓度荧光免疫测定法），该方法已被开发用于在活的患者肿瘤细胞、肿瘤细胞系或外周血细胞上快速筛选培养上清液。我们报告了一种乳腺癌抗体、五种结肠癌抗体、三种黑色素瘤抗体、三种肾瘤抗体和两种胰岛细胞癌抗体的研发情况。到目前为止，乳腺癌抗体能与75%的所检测乳腺癌结合，并且与五种结肠癌抗体一样，能与77 - 85%的结肠癌结合。所描述的黑色素瘤抗体能与90 - 100%的所检测黑色素瘤和前列腺癌发生反应。制备杂交瘤并筛选抗体以评估其潜在临床用途的整个过程耗时6到9个月完成，包括通过免疫组织化学检测正常组织反应性以及从腹水中生产克级量的单克隆抗体。
