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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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.
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