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A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins

Cochran, Siska, Li, Cai Ping, & Ferro, Vito (2009) A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins. Glycoconjugate Journal, 26(5), pp. 577-587.

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Abstract

A surface plasmon resonance-based solution affinity assay is described for measuring the Kd 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 Kd 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.

Impact and interest:

16 citations in Scopus
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16 citations in Web of Science®

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ID Code: 28440
Item Type: Journal Article
Keywords: Heparan sulfate-binding proteins, heparin, solution affinity assay, surface plasmon resonance
DOI: 10.1007/s10719-008-9210-0
ISSN: 0282-0080
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100) > Analytical Biochemistry (060101)
Divisions: Past > QUT Faculties & Divisions > Faculty of Science and Technology
Past > Schools > School of Physical & Chemical Sciences
Copyright Owner: Copyright 2008 Springer Science + Business Media, LLC
Copyright Statement: The original publication is available at SpringerLink http://www.springerlink.com
Deposited On: 09 Nov 2009 08:21
Last Modified: 29 Feb 2012 23:51

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