Barnstable C J, Bodmer W F, Brown G, Galfre G, Milstein C, Williams A F, Ziegler A
Cell. 1978 May;14(1):9-20. doi: 10.1016/0092-8674(78)90296-9.
Antibody-secreting hybrid cells have been derived from a fusion between mouse myeloma cells and spleen cells from a mouse immunized with membrane from human tonsil lymphocyte preparations. Hybrids secreting antibodies to cell surface antigens were detected by assaying culture supernatants for antibody binding to human tonsil cells. Six different antibodies (called W6/1, /28, /32, /34, /45 and /46 were analyzed. These were either against antigens of wide tissue distribution (W6/32, /34, and /46) or mainly on erythrocytes (W6/1 and W6/28). One of the anti-erythrocyte antibodies (W6/1) detected a polymorphic antigen, since blood group A1 and A2 erythrocytes were labeled while B and O were not. Antibodies W6/34, /45 and /46 were all against antigens which were mapped to the short arm of chromosome 11 by segregation analysis of mouse-human hybrids. Immunoprecipitation studies suggest that W6/45 antigen may be a protein of 16,000 dalton, apparent molecular weight, while W6/34 and /46 antigens could not be detected by this technique. Antibody W6/32 is against a determinant common to most, if not all, of the 43,000 dalton molecular weight chains of HLA-A, B and C antigens. This was established by somatic cell genetic techniques and by immunoprecipitation analysis. Tonsil leucocytes bound 370,000 W6/32 antibody molecules per cell at saturation. The hybrid myelomas W6/32 and W6/34 have been cloned, and both secrete an IgG2 antibody. W6/32 cells were grown in mice, and the serum of the tumor-bearing animals contained greater than 10 mg/ml of monoclonal antibody. The experiments established the usefulness of the bybrid myeloma technique in preparing monospecific antibodies against human cell surface antigens. In particular, this study highlights the possibilities not only of obtaining reagents for somatic cell genetics, but also of obtaining mouse antibodies detecting human antigenic polymorphisms.
分泌抗体的杂交细胞源自小鼠骨髓瘤细胞与用来自人扁桃体淋巴细胞制剂的膜免疫的小鼠脾细胞之间的融合。通过检测培养上清液与人扁桃体细胞的抗体结合来检测分泌针对细胞表面抗原的抗体的杂交细胞。分析了六种不同的抗体（称为W6/1、/28、/32、/34、/45和/46）。这些抗体要么针对广泛组织分布的抗原（W6/32、/34和/46），要么主要针对红细胞（W6/1和W6/28）。其中一种抗红细胞抗体（W6/1）检测到一种多态性抗原，因为A1和A2血型的红细胞被标记，而B和O血型的红细胞未被标记。抗体W6/34、/45和/46均针对通过小鼠-人杂交体的分离分析定位到11号染色体短臂上的抗原。免疫沉淀研究表明，W6/45抗原可能是一种表观分子量为16,000道尔顿的蛋白质，而W6/34和/46抗原用该技术无法检测到。抗体W6/32针对HLA-A、B和C抗原的43,000道尔顿分子量链中大多数（如果不是全部）共有的一个决定簇。这是通过体细胞遗传学技术和免疫沉淀分析确定的。扁桃体白细胞在饱和状态下每个细胞结合370,000个W6/32抗体分子。杂交骨髓瘤W6/32和W6/34已被克隆，两者均分泌IgG2抗体。W6/32细胞在小鼠体内生长，荷瘤动物的血清中含有大于10mg/ml的单克隆抗体。这些实验证实了杂交骨髓瘤技术在制备针对人细胞表面抗原的单特异性抗体方面的实用性。特别是，这项研究不仅突出了获得用于体细胞遗传学试剂的可能性，还突出了获得检测人抗原多态性的小鼠抗体的可能性。
