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Optimisation of the enzyme-linked lectin assay for enhanced glycoprotein and glycoconjugate analysis

Thompson, Roisin and Creavin, Aileen and O'Connell, Michael and O'Connor, Brendan 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

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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.

Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:Enzyme-linked lectin assay; Glycoprotein Analysis; Lectin; Glycoprotein; Blocking reagents; BSA; PVA
Subjects:Biological Sciences > Biotechnology
Biological Sciences > Biochemistry
Biological Sciences > Biosensors
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Research Initiatives 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 15 Feb 2017 16:57

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