Parallel analysis of glycoproteins using lectin-functionalised monoPLOT columns integrated in micro-fluidic chips
Vázquez, Mercedes and Nesterenko, Ekaterina and Clarke, Paul A. and Thompson, Roisin and O'Connor, Brendan and Brabazon, Dermot and Paull, Brett (2011) Parallel analysis of glycoproteins using lectin-functionalised monoPLOT columns integrated in micro-fluidic chips. In: Eurocarb 16, 3-7 Jul 2011, Sorrento , Italy.
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Glycoprotein isolation, purification and analysis is critical in the production of biologics. Therefore, there is a growing interest in new methods and techniques that allow ultra fast analytical characterisation and detection of glycoproteins and their glycoforms in the biological samples employed within the biopharmaceutical industry. In that context, micro-fluidic devices offer a great potential owing to the very fast analysis they allow, the low sample/reagent consumption, the disposability, and the ease of integration with other analytical systems, among others.
In this work, a micro-fluidic device integrating porous layer open tubular columns based on dimethacrylate monoliths (monoPLOT) and functionalised with Erythrina cristagalli lectin (ECL) were used for extraction of selected glycoproteins. Polymer monoliths are very versatile materials as they can be prepared with different porosities, pore sizes, and a wide variety of functionalities using many different precursors and chemistries. The monoPLOT columns were fabricated and functionalised off-chip, and then bonded to the channel walls before final channel lamination. In order to increase the monolith surface area, immobilisation of gold nano-particles (AuNPs) was carried out prior to covalent attachment of the ECL to the AuNPs. Detection was carried out with capacitively coupled contactless conductivity detection (C4D).
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