Solid-phase binding assay to study interaction of chemokines with glycosaminoglycans.

Publisher Summary This chapter develops a solid-phase binding assay for chemokine-glycosaminoglycan interactions. Optimization and validation studies were performed to recognize and avoid significant ligand depletion, retain a significant signal-to-noise ratio, explore the effects of buffer composition, and examine nonspecific binding. Results show that variations in immobilized heparin concentration and ionic strength will affect the IC values in competition experiments. The results from this type of assay are relative, rather than absolute measurements of the affinity between chemokines and immobilized heparin. Considering this limitation, the assay is suitable for making comparisons between the relative potencies of different glycosaminoglycans or other small molecule competitors, and it may be useful in identifying molecules that can selectively inhibit the interactions between chemokines and glycosaminoglycans. Molecules identified by this assay could be further tested in other systems to characterize their suitability as anti-inflammatory molecules.