Development and characterization of a high-throughput system for assessing cell-surface receptor-ligand engagement

Citation

Harbers, G. M.; Gamble, L. J.; Irwin, E. F.; Castner, D. G.; & Healy, K. E. (2005). Development and characterization of a high-throughput system for assessing cell-surface receptor-ligand engagement. Langmuir: the ACS journal of surfaces and colloids, 21(18), 8374-8384.

Abstract

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.

Keyword(s)

Cell Adhesion
Fibrinogen
Ligands
Molecular Structure
Peptides
Polystyrenes
Receptors, Cell Surface
Water

Reference Type

Journal Article

Secondary Title

Langmuir: the ACS journal of surfaces and colloids

Author(s)

Harbers, G. M.
Gamble, L. J.
Irwin, E. F.
Castner, D. G.
Healy, K. E.

Year Published

2005

Date Published

1125360000

Volume Number

21

Issue Number

18

Pages

8374-8384

DOI

10.1021/la050396y