Thompson, Roisin, Creavin, Aileen, O'Connell, Michael, O'Connor, Brendan ORCID: 0000-0002-8999-5184 and Clarke, Paul A. (2011) Optimisation of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis. Analytical Biochemistry, 413 (2). pp. 114-122. ISSN 1096-0309
Abstract
Lectin’s are proteins capable of recognising and binding to specific oligosaccharide tructures found on glycoproteins and other biomoloecules. As such they have found tility for glycoanalytical applications. One common difficulty encountered in the pplication of these proteins, particularly in multi-well plate assay formats known as Enzyme Linked Lectin Assays (ELLA’s), is in finding appropriate blocking solutions to prevent non-specific binding with plate surfaces. Many commonly used blocking agents contain carbohydrates and generate significant background signals in ELLA’s, limiting the utility of the assay.
In this study we examined the suitability of a range of blocking reagents, including rotein based, synthetic and commercially available carbohydrate free blocking eagents, for ELLA applications. Each blocking reagent was assessed against a panel f 19 commercially available biotinylated lectins exhibiting diverse structures and arbohydrate specificities. We identified the synthetic polymer Polyvinyl Alcohol PVA) as the best global blocking agent for performing ELLA’s. We ultimately present n ELLA methodology facilitating broad spectrum lectin analysis of glycoconjugates nd extending the utility of the ELLA.
Metadata
Item Type: | Article (Published) |
---|---|
Refereed: | Yes |
Uncontrolled Keywords: | Enzyme-linked lectin assay; Glycoprotein Analysis; Lectin; Glycoprotein; Blocking reagents; BSA; PVA |
Subjects: | Biological Sciences > Biotechnology Humanities > Biological Sciences > Biotechnology Biological Sciences > Biochemistry Humanities > Biological Sciences > Biochemistry Biological Sciences > Biosensors Humanities > Biological Sciences > Biosensors |
DCU Faculties and Centres: | DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology Research Institutes and Centres > Irish Separation Science Cluster (ISSC) |
Publisher: | Elsevier |
Official URL: | http://dx.doi.org/10.1016/j.ab.2011.02.013 |
Copyright Information: | © 2011 Elsevier |
Use License: | This item is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 3.0 License. View License |
Funders: | Science Foundation Ireland |
ID Code: | 17772 |
Deposited On: | 14 Feb 2013 09:19 by Brendan O'connor . Last Modified 03 Oct 2018 14:58 |
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