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Localized Cell-Surface Sampling of a Secreted Factor Using Cell-Targeting Beads.
- Source :
-
Analytical chemistry [Anal Chem] 2020 Oct 20; Vol. 92 (20), pp. 13634-13640. Date of Electronic Publication: 2020 Oct 02. - Publication Year :
- 2020
-
Abstract
- Intercellular communication through the secretion of soluble factors plays a vital role in a wide range of biological processes (e.g., homeostasis, immune response), yet identification and quantification of many of these factors can be challenging due to their degradation or sequestration in cell culture media prior to analysis. Here, we present a customizable bead-based system capable of simultaneously binding to live cells (through antibody-mediated cell tethering) and capturing cell-secreted molecules. Our functionalized beads capture secreted molecules (e.g., hepatocyte growth factor secreted by fibroblasts) that are diminished when sampled via traditional supernatant analysis techniques ( p < 0.05), effectively rescuing a reduced signal in the presence of neutralizing components in the cell culture media. Our system enables capture and analysis of molecules integral to chemical communication that would otherwise be markedly decreased prior to analysis.
- Subjects :
- Antibodies immunology
Antigens, Surface analysis
Antigens, Surface immunology
Cell Line
Culture Media chemistry
Fibroblasts cytology
Fibroblasts metabolism
Fluoresceins chemistry
Hepatocyte Growth Factor immunology
Hepatocyte Growth Factor metabolism
Human Umbilical Vein Endothelial Cells
Humans
Immunoassay methods
Thy-1 Antigens analysis
Thy-1 Antigens immunology
Thy-1 Antigens metabolism
Antigens, Surface metabolism
Hepatocyte Growth Factor analysis
Microspheres
Subjects
Details
- Language :
- English
- ISSN :
- 1520-6882
- Volume :
- 92
- Issue :
- 20
- Database :
- MEDLINE
- Journal :
- Analytical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 32941013
- Full Text :
- https://doi.org/10.1021/acs.analchem.0c02578