Comparison of results using the gel microdrop cytokine secretion assay with ELISPOT and intracellular cytokine staining assay

被引：10

作者：

Atochina, O ^{[1
]}

Mylvaganam, R ^{[1
]}

Akselband, Y ^{[1
]}

McGrath, P ^{[1
]}

机构：

[1] One Cell Syst Inc, Cambridge, MA USA

来源：

CYTOKINE | 2004年 / 27卷 / 4-5期

关键词：

cytokines; ELISPOT; flow cytometry;

D O I：

10.1016/j.cyto.2004.04.003

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The gel microdrop (GMD) secretion assay involves encapsulating single cells in a biotinylated agarose matrix, addition of a streptavidin bridge, diffusion of a biotinylated capture antibody, and detection of secreted molecules using a fluorescently labeled reporter antibody. Using flow cytometry, encapsulated cells can be analyzed or recovered based on cell type and secretory profile. Using murine Th2 cell line D10.G4.1 as a model, we recently demonstrated the feasibility of using the GMD cytokine secretion assay combined with flow cytometry to detect IL-4-producing cells after stimulation with the mitogen, Con A. In addition, subpopulations of encapsulated cells secreting IL-4 were simultaneously characterized by immunophenotyping. We found good correlation between results using the GMD cytokine secretion assay and results with the standard ELISPOT and intracellular cytokine (ICC) assays. The GMD cytokine secretion assay permits simultaneous detection of secreted cytokine and determination of cell surface phenotype on viable, single cells. Moreover, using fluorescence activated cell sorting (FACS), secreting cells of interest can be sorted, recovered, and cultured for further studies. (C) 2004 Elsevier Ltd. All rights reserved.

引用

页码：120 / 128

页数：9

共 22 条

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