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Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

O'Shea, Vincent and Clarke, Paul A. and O'Connor, Brendan and Paull, Brett (2007) Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection. Journal of Separation Science, 30 (17). pp. 3060-3068. ISSN 1615-9306

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Abstract

Capacitively coupled contactless conductivity detection (C4D) is presented as a novel and versatile means of visualising discrete zones of charged functional groups grafted onto polymer based monoliths. Monoliths were formed within 100 μm UV transparent fused silica capillaries and photografting methods were subsequently used to graft a charged functional monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) onto discrete regions of the “generic” monolith using a photomask. Post-modification monolith evaluation involves scanning the C4D detector along the length of the monolith to obtain a profile of the exact spatial location of grafted charged functionalities with millimetre accuracy. The methodology was extended to the visualisation of several zones of immobilised protein (bovine serum albumin) using photografted azlactone groups to enable covalent attachment of the protein to the monolith at precise locations along its length. In addition, the extent of non-specific binding of protein to the ungrafted regions of the monolith due to hydrophobic interactions could be monitored as an increase in background conductivity of the stationary phase. Finally, the technique was cross-validated using fluorescence microscopy by immobilising green fluorescent protein (GFP) in discrete zones and comparing the profiles obtained using both complementary techniques.

Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:Polymer monolithic phase; Photografted protein; Contactless conductivity detection
Subjects:Biological Sciences > Biotechnology
Physical Sciences > Analytical chemistry
Biological Sciences > Biochemistry
Biological Sciences > Molecular biology
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Chemical Sciences
DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Research Initiatives and Centres > Irish Separation Science Cluster (ISSC)
Publisher:John Wiley & Sons
Official URL:http://dx.doi.org/10.1002/jssc.200700365
Copyright Information:© 2007 Wiley The definitive version is available at www3.interscience.wiley.com
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 3.0 License. View License
Funders:Science Foundation Ireland, Enterprise Ireland
ID Code:17803
Deposited On:28 Feb 2013 14:02 by Brendan O'Connor. Last Modified 15 Feb 2017 17:00

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