Abstract
Affinity chromatography is a valuable method to purify and concentrate minute amount of proteins. Monoliths with epoxy groups for affinity immobilization were prepared by direct in-situ photopolymerization of glycidyl methacrylate and ethylene glycol dimethacrylate in porogenic solvents consisting of 1-dodecanol and cyclohexanol. By integrating affinity monoliths onto a microfluidic system, targeted biomolecules can be captured and retained on affinity column, while other biomolecules having no specific interactions toward the immobilized ligands flow through the microchannel. Therefore, proteins which remain on the affinity column are purified and concentrated, and then eluted by appropriate solutions and finally, separated by microchip capillary electrophoresis. This integrated microfluidic device has been applied to the purification and separation of specific proteins (FITC-labeled human serum albumin and IgG) in a mixture.
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References
Vickova M, Stettler A, Schwarz M (2006) Microchip affinity capillary electrophoresis: applications and recent advances. J Liq Chromatogr Relat Technol 29:1047–1076
Schou C, Heegaard NHH (2006) Recent applications of affinity interactions in capillary electrophoresis. Electrophoresis 27:44–59
Yang W, Sun X, Wang H et al (2009) Integrated microfluidic device for serum biomarker quantitation using either standard addition or a calibration curve. Anal Chem 81:8230–8235
Gong M, Wehmeyer KR, Limbach PA et al (2006) On-line sample preconcentration using field-amplified stacking injection in microchip capillary electrophoresis. Anal Chem 78:3730–3737
Song S, Singh AK, Shepodd TJ et al (2004) Microchip dialysis of proteins using in situ photopatterned nanoporous polymer membranes. Anal Chem 76:2367–2373
Wolfe KA, Breadmore MC, Ferrance JP et al (2002) Toward a microchip-based solid-phase extraction method for isolation of nucleic acids. Electrophoresis 23:727–733
Huynh BH, Fogarty BA, Martin RS et al (2004) On-Line coupling of microdialysis sampling with microchip-based capillary electrophoresis. Anal Chem 76:6440–6447
Roddy ES, Price M, Ewing AG (2003) Continuous monitoring of a restriction enzyme digest of DNA on a microchip with automated capillary sample introduction. Anal Chem 75:3704–3711
Currivan S, Jandera P (2014) Post-polymerization modifications of polymeric monolithic columns. Chromatography 1:24–53
Trauner A, Mark HB, Williams HD (2011) Isolation of bacterial ribosomes with monolith chromatography. PLoS One 4:e16273
Yu C, Davey MH, Svec F et al (2001) Monolithic porous polymer for on-chip solid-phase extraction and preconcentration prepared by photoinitiated in situ polymerization within a microfluidic device. Anal Chem 73:5088–5096
Breadmore MC, Wolfe KA, Arcibal IG et al (2003) Microchip-based purification of DNA from biological samples. Anal Chem 75:1880–1886
Sun X, Yang W, Pan T et al (2008) Affinity monolith-integrated poly(methyl methacrylate) microchips for on-line protein extraction and capillary electrophoresis. Anal Chem 80:5126–5130
Hahn R, Podgornik A, Merhar M et al (2001) Affinity monoliths generated by in situ polymerization of the ligand. Anal Chem 73:5126–5132
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Gao, C., Sun, X., Wang, H., Qiao, W., Hu, B. (2016). Affinity Monolith-Integrated Microchips for Protein Purification and Concentration. In: Tran, N., Taverna, M. (eds) Capillary Electrophoresis of Proteins and Peptides. Methods in Molecular Biology, vol 1466. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4014-1_7
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DOI: https://doi.org/10.1007/978-1-4939-4014-1_7
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