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通过细胞分选分离具有预定义可变表面免疫球蛋白的骨髓瘤变体。

Isolation of myeloma variants with predefined variant surface immunoglobulin by cell sorting.

作者信息

Liesegang B, Radbruch A, Rajewsky K

出版信息

Proc Natl Acad Sci U S A. 1978 Aug;75(8):3901-5. doi: 10.1073/pnas.75.8.3901.

Abstract

We describe a procedure for the isolation of somatic cell variants in which gene products are expressed on the cell surface that are not expressed in the wild type. Cloned cells of the myeloma line MPC 11, which expresses an IgG2b protein, were incubated with an antiserum specific for IgGI and IgG2a. Cells reacting with this antiserum were stained with a fluorescent anti-antiserum and enriched in three cycles of sorting in the fluorescence-activated cell sorter and subsequent growth in vitro. From the enriched population two variants were isolated by cloning in soft agar. One of them expressed a variant immunoglobulin that types serologically as an IgG2a but whose variable portion was idiotypically related to that of the MPC 11 wild-type protein.

摘要

我们描述了一种分离体细胞变体的方法,其中基因产物在细胞表面表达,而在野生型中不表达。表达IgG2b蛋白的骨髓瘤细胞系MPC 11的克隆细胞与针对IgG1和IgG2a的抗血清一起孵育。与该抗血清反应的细胞用荧光抗抗血清染色,并在荧光激活细胞分选仪中进行三个分选循环,随后在体外培养以富集。通过在软琼脂中克隆,从富集群体中分离出两个变体。其中一个表达一种变体免疫球蛋白,血清学类型为IgG2a,但其可变部分在独特型上与MPC 11野生型蛋白相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2890/392896/a0d3393499fc/pnas00020-0373-a.jpg

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