Bumol T F, Chee D O, Reisfeld R A
Hybridoma. 1982;1(3):283-92. doi: 10.1089/hyb.1.1982.1.283.
Immunoprecipitation studies with application of monoclonal antibody F11 originally made to a partially purified spent medium antigen of melanoma cells, made it possible to delineate the molecular profiles of both the cell associated and spent medium antigens recognized on melanoma cells intrinsically labeled with glycoprotein precursors. F11 distinguishes a glycoprotein of Mr 100,000 (100 K) in the spent media of melanoma cells while a parallel analysis of detergent lysates of cells reveals a pattern of three glycoproteins of Mr 75, 77, and 100 K. Pulse-chase analysis of the biosynthesis of these antigens indicated that F11 first recognizes the 75 and 77 K antigens in the absence of a 100K component suggesting strongly that these molecules contain an antigenic site recognized by F11. The 100 K antigen appears later in the pulse-chase analysis with kinetics that suggest some of the 75 and 77 K antigens are biosynthetic precursors of the 100 K antigen. This molecule is ultimately secreted into the extracellular media and appears to be a sialoglycoprotein judging from its sensitivity to neuraminidase. A cross-reactive species with an approximate Mr 90 K is also recognized by F11 in indirect immunoprecipitation analysis of spent media from a neuroblastoma cell line indicating that a common antigenic site exists on this secreted but structurally different neuroblastoma antigen. Thus, a combination of immunochemical and biosynthetic analyses of cell-associated and secreted antigens recognized by monoclonal antibody F11 demonstrate such molecules can differ structurally when isolated from the same or different tumor cells. These findings indicate the necessity to establish molecular profiles of melanoma-associated glycoprotein antigens recognized by monoclonal antibodies to characterize and define their potential biological functions within tumor cells.
使用最初针对黑色素瘤细胞部分纯化的废培养基抗原制备的单克隆抗体F11进行免疫沉淀研究，使得描绘在糖蛋白前体内在标记的黑色素瘤细胞上识别的细胞相关抗原和废培养基抗原的分子谱成为可能。F11在黑色素瘤细胞的废培养基中识别出一种分子量为100,000（100K）的糖蛋白，而对细胞去污剂裂解物的平行分析揭示了分子量为75K、77K和100K的三种糖蛋白模式。对这些抗原生物合成的脉冲追踪分析表明，在没有100K成分的情况下，F11首先识别75K和77K抗原，这强烈表明这些分子含有F11识别的抗原位点。在脉冲追踪分析中，100K抗原稍后出现，其动力学表明一些75K和77K抗原是100K抗原的生物合成前体。这种分子最终分泌到细胞外培养基中，从其对神经氨酸酶的敏感性判断，它似乎是一种唾液酸糖蛋白。在对神经母细胞瘤细胞系废培养基的间接免疫沉淀分析中，F11还识别出一种分子量约为90K的交叉反应物种，这表明在这种分泌但结构不同的神经母细胞瘤抗原上存在一个共同的抗原位点。因此，对单克隆抗体F11识别的细胞相关和分泌抗原进行免疫化学和生物合成分析相结合，表明从相同或不同肿瘤细胞中分离时，这些分子在结构上可能不同。这些发现表明有必要建立单克隆抗体识别的黑色素瘤相关糖蛋白抗原的分子谱，以表征和定义它们在肿瘤细胞内的潜在生物学功能。
