Winkelhake J L, Buckmire F L
Cancer Res. 1977 Apr;37(4):1197-204.
A radioimmune assay for the antitumor agent, macromomycin, using purified, radioiodine-labeled macromomycin and antisera raised in rabbits against a carbodiimide-catalyzed macromycin-Limulus polyphemus hemocyanin complex has been developed. Radiolabeled macromomycin was prepared by direct iodination of the polypeptide antibiotic with the use of iodine monochloride or solid-state lactoperoxidase. Antibody-bound drug was isolated from free macromomycin with dextran-coated, activated charcoal. The standard curve of the sequential saturation assay was linear on a logit-log plot and indicated a lower limit of sensitivity of approximately 100 pg macromomycin. The radioimmune assay was suitable for measuring macromomycin in the presence of other antitumor drugs, and detection of macromomycin was quantitative when it was added to normal human serum or urine. Drug binding to melanoma and mammary carcinoma cell surfaces could be inhibited by preincubating macromomycin with affinity-purified antimacromomycin antibodies. However, once the drug was bound to cell surfaces, addition of antimacromomycin antibodies did not result in removal of the drug from cell surfaces or in reversal of macromomycin-induced inhibition of thymidine incorporation into cellular DNA. Antimacromovide useful tools for developing pharmacokinetic and toxicity studies of macromomycin, as well as for analyzing the mechanism(s) of action of the drug.
已开发出一种用于抗肿瘤药物大分子霉素的放射免疫测定法，该方法使用纯化的、放射性碘标记的大分子霉素以及在兔体内产生的针对碳二亚胺催化的大分子霉素-鲎血蓝蛋白复合物的抗血清。放射性标记的大分子霉素通过使用一氯化碘或固态乳过氧化物酶对多肽抗生素进行直接碘化来制备。用葡聚糖包被的活性炭从游离的大分子霉素中分离出与抗体结合的药物。序列饱和测定法的标准曲线在logit-log图上呈线性，表明灵敏度下限约为100 pg大分子霉素。该放射免疫测定法适用于在存在其他抗肿瘤药物的情况下测定大分子霉素，并且当将大分子霉素添加到正常人血清或尿液中时，其检测是定量的。通过将大分子霉素与亲和纯化的抗大分子霉素抗体预孵育，可以抑制药物与黑色素瘤和乳腺癌细胞表面的结合。然而，一旦药物与细胞表面结合，添加抗大分子霉素抗体并不会导致药物从细胞表面去除，也不会导致大分子霉素诱导的胸腺嘧啶核苷掺入细胞DNA的抑制作用逆转。抗大分子霉素抗体是开展大分子霉素药代动力学和毒性研究以及分析该药物作用机制的有用工具。
