Improved cell line development by a high throughput affinity capture surface display technique to select for high secretors

被引:48
作者
Holmes, P [1 ]
Al-Rubeai, M [1 ]
机构
[1] Univ Birmingham, Sch Chem Engn, Anim Cell Technol Grp, Birmingham B15 2TT, W Midlands, England
关键词
affinity capture surface display; secretion assay; cell selection; cell enrichment; high throughput screening; FACS;
D O I
10.1016/S0022-1759(99)00181-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel process is described which permits rapid and objective selection of rare cells from a heterogeneous population based on quantity of secreted target protein. The process involves construction of an immobilised affinity surface display matrix that specifically binds secreted target product which is then detected using a fluorescent labelled ligand. Cells with the highest fluorescence can then be sorted using conventional flow cytometric technology. Overall, the whole process can be completed in less than 4 h during which time in the region of five million cells can be analysed. Cells are rapidly selected for in a quantitative manner compared to traditional methods which can tale several months and have a reduced probability of finding low abundance high secretors due to practical limitations imposed on the number of cells which can be screened. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:141 / 147
页数:7
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