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Pipette-tip selective extraction of glycoproteins with lectin modified gold nano-particles on a polymer monolithic phase

Citation

Alwael, H and Connolly, D and Clarke, P and Thompson, R and Twamley, B and O'Connor, B and Paull, B, Pipette-tip selective extraction of glycoproteins with lectin modified gold nano-particles on a polymer monolithic phase, Analyst, 136, (12) pp. 2619-2628. ISSN 0003-2654 (2011) [Refereed Article]


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Copyright Statement

Copyright 2011 Royal Society of Chemistry

DOI: doi:10.1039/c1an15137a

Abstract

The in situ preparation of ethylene dimethacrylate porous polymer monoliths within 20 mu L polypropylene pipette tips, bound via surface grafted methacrylate anchor sites, is reported. Gold nano-particles (AuNPs) were immobilised onto the monolith pore surface utilising azlactone chemistry and coverage verified using field emission scanning electron microscopy. Erythrina cristagalli lectin (ECL) was immobilised upon the attached AuNPs via a bio-functional linker. The ECL-modified tip was successfully applied for the enrichment of galactosylated protein (desialylated transferrin) versus a non-galactosylated protein (ribonuclease B) due to the specificity of ECL. Reversed-phase capillary HPLC was used to validate the efficiency and selectivity of the developed micro-extraction phase which resulted in an increase in extraction recovery of similar to 95% due to the AuNP enhanced surface area. Further specificity of the ECL-modified tip was demonstrated with a complex mixture of non-glycosylated and glycosylated proteins with differing terminal sugar structures. Finally, the lectin affinity phase was applied to a galactosylated glycoproteins spiked Escherichia coli cell lysate to successfully demonstrate matrix tolerance.

Item Details

Item Type:Refereed Article
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Separation Science
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Chemical Sciences
Author:Paull, B (Professor Brett Paull)
ID Code:72674
Year Published:2011
Web of Science® Times Cited:38
Deposited By:Chemistry
Deposited On:2011-08-30
Last Modified:2017-10-29
Downloads:1 View Download Statistics

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