19034645 |
Cochran S, Li CP, Ferro V: A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins. Glycoconj J. 2009 Jul;26(5):577-87. Epub 2008 Nov 26. A surface plasmon resonance-based solution affinity assay is described for measuring the K (d) of binding of heparin/heparan sulfate-binding proteins with a variety of ligands. The assay involves the passage of a pre-equilibrated solution of protein and ligand over a sensor chip onto which heparin has been immobilised. Heparin sensor chips prepared by four different methods, including biotin-streptavidin affinity capture and direct covalent attachment to the chip surface, were successfully used in the assay and gave similar K (d) values. The assay is applicable to a wide variety of heparin/HS-binding proteins of diverse structure and function (e.g., FGF-1, FGF-2, VEGF, IL-8, MCP-2, ATIII, PF4) and to ligands of varying molecular weight and degree of sulfation (e.g., heparin, PI-88, sucrose octasulfate, naphthalene trisulfonate) and is thus well suited for the rapid screening of ligands in drug discovery applications. |
31(0,1,1,1) |