Isolation of rare isotype switch variants in hybridoma cell lines using an agarose gel microdrop-based protein secretion assay

被引:12
作者
Akselband, Y [1 ]
Moen, PT [1 ]
McGrath, P [1 ]
机构
[1] One Cell Syst Inc, Cambridge, MA 02139 USA
关键词
D O I
10.1089/154065803770380977
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Using gel microdrop (GMD) encapsulation technology and fluorescence-activated cell storing (FACS), we have developed a rapid, sensitive, and reliable method for discriminating and recovering rare isotypic switch variants in hybridoma cell lines. Using the GMD-based IgSwitch(TM) assay, a novel approach for isolating subpopulations of IgG-secreting hybridoma cells present at a frequency of similar to1-10 in 10(6), We successfully isolated spontaneous and in vitro-induced isotypic switch variants in less than half the time required for conventional sublining. The effectiveness and specificity of the assay are demonstrated.
引用
收藏
页码:619 / 626
页数:8
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