Novel approach to construction of human "myeloma analogues" for production of human monoclonal antibodies.
Novel approach to construction of human "myeloma analogues" for production of human monoclonal antibodies.
复制标题
构建人类“骨髓瘤类似物”以生产人类单克隆抗体的新方法。
作者：
Posner,MR;Schlossman,SF;Lazarus,H
人单克隆抗体的生产受到缺乏人骨髓瘤细胞系的阻碍，所述细胞系容易生长、有效融合、容易克隆并且连续分泌大量抗体。通过将HAT敏感的非分泌性人骨髓瘤细胞系LSM 1.2与来自浆细胞白血病患者的细胞融合，构建了细胞系HM 2.0。与不能支持免疫球蛋白分泌的LSM 1.2形成鲜明对比的是，HM 2.0与来自脾或外周血的细胞的融合通常导致针对肺炎球菌多糖和破伤风类毒素的抗体的分泌。HM2.0的融合效率大于1/1.2 × 103，分泌特异性抗体的杂交细胞数大于1/1.1 × 105。这是对我们先前描述的人“骨髓瘤类似物”LSM 2.7的改进，所述人“骨髓瘤类似物”LSM 2.7通过将HAT敏感的非分泌性人骨髓瘤细胞系LSM 1.1与来自正常供体的细胞以及所有先前描述的人淋巴母细胞和骨髓瘤细胞系融合而衍生。这些结果表明，体细胞杂交可用于修饰现有的细胞系，以产生具有生产人单克隆抗体所需属性的“新”细胞系。
The production of human monoclonal antibodies has been impeded by the lack of human myeloma cell lines which grow easily, fuse efficiently, clone readily, and continuously secrete large amounts of antibody. A cell line, HM 2.0, was constructed by fusing a HAT-sensitive, nonsecreting, human myeloma cell line, LSM 1.2, with cells from a patient with plasma cell leukemia. In marked contrast to LSM 1.2, which could not support the secretion of immunoglobulin, fusion of HM 2.0 with cells from spleen or peripheral blood routinely resulted in the secretion of antibody to pneumococcal polysaccharides and tetanus toxoid. The fusion efficiency of HM 2.0, as measured by growth of colonies, was greater than 1 per 1.2 x 103peripheral blood mononuclear cells and the number of hybrids secreting specific antibody was greater than 1 per 1.1 x 105mononuclear cells from immunized individuals. This is an improvement over our previously described human "myeloma analogue" LSM 2.7, derived by fusion of a HAT-sensitive, nonsecreting human myeloma cell line, LSM 1.1, with cells from a normal donor, as well as all previously described human lymphoblastoid and myeloma cell lines. These results demonstrate that somatic cell hybridization can be used to modify an existing cell line in such a manner as to yield a "new" cell line with the attributes necessary for the production of human monoclonal antibodies.