SECRETION CAPTURE AND REPORT WEB - USE OF AFFINITY DERIVATIZED AGAROSE MICRODROPLETS FOR THE SELECTION OF HYBRIDOMA CELLS

被引：38

作者：

GRAY, F ^{[1
]}

KENNEY, JS ^{[1
]}

DUNNE, JF ^{[1
]}

机构：

[1] SYNTEX INC,DISCOVERY RES,PALO ALTO,CA 94304

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1995年 / 182卷 / 02期

关键词：

MONOCLONAL ANTIBODY; ISOLATION AND PURIFICATION; SECRETION; BIOSYNTHESIS; FLOW CYTOMETRY; MICRODROP;

D O I：

10.1016/0022-1759(94)00319-R

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel assay is described which allows the entrapment and detection of the immunoglobulin secreted from individual viable hybridoma cells using a secretion capture and reporter web (SCRW). By encapsulating the cells in agarose microdroplets which have been derivatized to create a fluorescent antigen-specific sandwich assay, flow cytometry can be used to identify and sort productive cells from a heterogeneous population. Using agarase, the cells can be recovered from the microdroplets and clonally expanded after selection. The assay has been used to redone rare secretors from hybridoma cultures and to enhance the production of cultures with poor producers. The assay is easily generalized for the detection of any secreted protein for which specific antibodies or other ligands are available.

引用

页码：155 / 163

页数：9