Manabe Y, Tsubota T, Haruta Y, Kataoka K, Okazaki M, Haisa S, Nakamura K, Kimura I
Acta Med Okayama. 1984 Apr;38(2):117-24. doi: 10.18926/AMO/30332.
Macromomycin (MCR), an unique membrane-reactive anticancer antibiotic, was incubated with murine monoclonal anti-HLA IgG1 antibody (H-1) in the presence of carbodiimide. The resulting mixture was fractionated with a Sephadex G-200 column. The first and second fractions were shown to contain MCR-(H-1) conjugate by the elution profile, as well as by the Sarcina lutea growth inhibition assay and Ouchterlony double-diffusion method. A membrane immunofluorescence test with anti-MCR and anti-mouse IgG antibodies demonstrated specific localization of MCR-(H-1) on the surface of HLA-bearing NALL -1 cells. MCR-(H-1) inhibited the growth of HLA-lacking NS-1 cells statistically less effectively than MCR alone (p less than 0.01). On the other hand, the conjugate and free MCR equally inhibited the growth and 3H-TdR incorporation of HLA-bearing NALL -1 cells. These results indicate that the antibody-bound MCR retained both MCR and antibody activities, and thus exerted antibody-targeting MCR cytotoxicity in vitro.
巨霉素（MCR）是一种独特的具有膜反应性的抗癌抗生素，在碳二亚胺存在的情况下，将其与鼠单克隆抗HLA IgG1抗体（H-1）一起孵育。所得混合物用Sephadex G - 200柱进行分级分离。通过洗脱曲线以及藤黄八叠球菌生长抑制试验和双向免疫扩散法表明，第一和第二级分含有MCR -（H-1）缀合物。用抗MCR和抗小鼠IgG抗体进行的膜免疫荧光试验表明，MCR -（H-1）特异性定位于携带HLA的NALL -1细胞表面。统计学上，MCR -（H-1）抑制缺乏HLA的NS-1细胞生长的效果比单独使用MCR差（p小于0.01）。另一方面，缀合物和游离MCR对携带HLA的NALL -1细胞的生长和3H - TdR掺入具有同等的抑制作用。这些结果表明，抗体结合的MCR保留了MCR和抗体活性，因此在体外发挥了抗体靶向的MCR细胞毒性。
