Galfrè G, Milstein C
Immunology. 1982 Jan;45(1):125-8.
Hybrids between human spleen cells and the non-secretor NSO mouse myeloma, and also between the rat non-secretor line YB2/O and human peripheral blood cells were prepared. After a month in culture very few hybrids retained the ability to secrete the human kappa light chain. From these, clones could be derived which remained stable over several months of continuous culture. On incorporating [3H]-Leu into the culture medium the cells secrete large amounts of radioactive light chain. It is shown that, even without dialysis, the purity of the preparation is sufficient for an automatic N-terminal sequence analysis at the radioactive level. From the pattern of distribution of leucine in the first twenty-two amino acid residues, it is possible to assign the synthesized light chains to one of the four established human subgroups. The method permits a fast and simple classification of human light chains secreted by hybrid myelomas. Although tested with rodent x human hybrids, we see no reason why the method could not equally apply to human x human hybrids.
制备了人脾细胞与非分泌型NSO小鼠骨髓瘤细胞的杂种细胞,以及大鼠非分泌型细胞系YB2/O与人外周血细胞的杂种细胞。培养一个月后,只有极少数杂种细胞仍保留分泌人κ轻链的能力。从这些细胞中可以获得在连续培养数月中仍保持稳定的克隆。将[3H]-亮氨酸掺入培养基后,细胞分泌大量放射性轻链。结果表明,即使不进行透析,制备物的纯度也足以在放射性水平上进行自动N端序列分析。根据前二十二个氨基酸残基中亮氨酸的分布模式,可以将合成的轻链归为已确定的四个人类亚群之一。该方法可快速、简单地对杂交骨髓瘤分泌的人轻链进行分类。尽管该方法是用啮齿动物与人的杂种细胞进行测试的,但我们认为该方法同样适用于人与人的杂种细胞没有理由不成立。