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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
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 |
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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 |
UTAS Author: | Paull, B (Professor Brett Paull) |
ID Code: | 72674 |
Year Published: | 2011 |
Web of Science® Times Cited: | 65 |
Deposited By: | Chemistry |
Deposited On: | 2011-08-30 |
Last Modified: | 2017-10-29 |
Downloads: | 1 View Download Statistics |
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