This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (4) Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Alban, S. Articles by Gastpar, R. Search for Related Content PubMed PubMed Citation Articles by Alban, S. Articles by Gastpar, R. Pubmed/NCBI databases Substance via MeSH Social Bookmarking                         What's this?

Development of SPC-ELISA: A New Assay Principle for the Study of Sulfated Polysaccharide-Protein Interactions

Institute of Pharmacy, University of Regensburg, Regensburg, Germany

Robert Gastpar

Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany

A sulfated polysaccharide-coating enzyme-linked immunosorbent assay (SPC-ELISA), a new screening assay for the study of interactions between sulfated polysaccharides and proteins, has been developed. Fibrinogen was used as representative for the protein. A microplate is coated with the sulfated polysaccharide to be tested and then incubated with various concentrations of fibrinogen. The bound fibrinogen is quantified by ELISA technique. The assay has been optimized with respect to coating procedure, incubation times, antibody concentrations, and detection conditions. Its capacity was demonstrated using three different sulfated polysaccharides: heparin, a sulfated glucuronogalactan extracted from a red algae, and a semisynthetic xanthan sulfate. Furthermore, the fibrinogen binding of semisynthetic laminarin sulfates with different degrees of sulfation showed good correlation with their anticoagulant effect as measured by fibrinogen clotting time. The intraassay as well as the interassay variations were lower than 8%. The binding properties observed in the SPC-ELISA correlated well with those found utilizing conventional gel permeation chromatography and fibrinogen affinity gel electrophoresis. Compared to these methods, the SPC-ELISA has several advantages: It is more rapid and far easier to perform, allows high throughput screening, and is suitable for automation. Furthermore, it is inexpensive, highly sensitive, and reproducible and has no special equipment requirements. Finally, the method represents the basis for multiple variations with regard to the target proteins as well as the detection methods.

Journal of Biomolecular Screening, Vol. 6, No. 6, 393-400 (2001)
DOI: 10.1177/108705710100600605


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				A. Basu, T. Kanda, A. Beyene, K. Saito, K. Meyer, and R. Ray Sulfated Homologues of Heparin Inhibit Hepatitis C Virus Entry into Mammalian Cells J. Virol., April 15, 2007; 81(8): 3933 - 3941. [Abstract] [Full Text] [PDF]

				E. J. Joo, G. B. ten Dam, T. H. van Kuppevelt, T. Toida, R. J. Linhardt, and Y. S. Kim Nucleolin: acharan sulfate-binding protein on the surface of cancer cells Glycobiology, January 1, 2005; 15(1): 1 - 9. [Abstract] [Full Text] [PDF]