Development and characterization of a high-throughput system for assessing cell-surface receptor-ligand engagement
| Autor: | Gamble Lj, Kevin E. Healy, Gregory M. Harbers, David G. Castner, Elizabeth F. Irwin |
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| Rok vydání: | 2005 |
| Předmět: |
Molecular Structure
Ligand Fibrinogen Water Receptors Cell Surface Surfaces and Interfaces Adhesion Condensed Matter Physics Ligands Contact angle chemistry.chemical_compound chemistry Polymer chemistry PEG ratio Electrochemistry Cell Adhesion Polystyrenes General Materials Science Interpenetrating polymer network Polystyrene Peptides Spectroscopy Acrylic acid Protein adsorption |
| Zdroj: | Langmuir : the ACS journal of surfaces and colloids. 21(18) |
| ISSN: | 0743-7463 |
| Popis: | A nonfouling interfacial interpenetrating polymer network (IPN) of poly(acrylamide-co-ethylene glycol/acrylic acid) [p(AAm-co-EG/AAc)] was grafted to polystyrene for use as a novel platform for the development of high-throughput assays for screening of specific bimolecular interactions (i.e., receptor-ligand engagement). For the development of the IPN, a water-soluble hydrogen-abstracting photoinitiator was investigated: (4-benzoylbenzyl)trimethylammonium chloride. IPN-modified polystyrene surfaces were characterized using XPS, contact angle goniometry, and protein adsorption analysis. These IPN surfaces minimized fibrinogen adsorption compared to tissue culture polystyrene (96% reduction), prevented mammalian cell adhesion, and served as nonfouling surfaces to graft biological ligands. For bimolecular interaction studies, a model peptide ligand from bone sialoprotein (Ac-CGGNGEPRGDTYRAY-NH(2)) was grafted to p(AAm-co-EG/AAc) via a 3400 M(w) linear pEG spacer. Ligand density measurements, cell culture, and a centrifugal adhesion assay were used to study cell adhesion to peptide-modified IPNs (i.e., receptor-ligand engagement). Ligand density (Gamma) was controllable from approximately 1 to 20 pmol/cm(2) by modulating the peptide input concentration (0.02-20 microM). Cell adhesion was directly dependent on the ligand density. This technology creates a powerful high-throughput system to simultaneously probe a myriad of cell-surface receptor-ligand interactions. |
| Databáze: | OpenAIRE |
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