Affinity separations using microfabricated microfluidic devices:In situ photopolymerization and use in protein separations
- 60 Downloads
The use of microfabricated microfluidic devices offers significant advantages over current technologies including fast analysis time and small reagent requirements. In the context of proteomic research, the possibility of using affinity-based separations for prefractionation of samples using microfluidic devices has significant potential. We demonstrate the use of microscale devices to achieve affinity separations of proteins using a device fabricated from borosilicate glass wafers. Photolithography and wet etching are used to pattern individual glass wafers and the wafers are fusion bonded at 650°C to obtain enclosed channels. A polymer has been successfully polymerizedin situ and used either as a frit for packing beads or, when derivatized with Cibacron Blue 3GA, as a separation matrix. Both of these technologies are based onin situ UV photopolymerization of glycidyl methacrylate (GMA) and trimethylolpropane trimethacrylate (TRIM) in channels.
Keywordsaffinity separation microscale devices nanobiotechnology
Unable to display preview. Download preview PDF.
- Liu, H., D. Lin, and J. H. Yates, III (2002) Review: Multidimensional separations for protein/peptide analysis in the post-genomic era.Bio Techniques 32: 898–911.Google Scholar
- Gavin, A. C., M. Bosche, R. Krause, P. Grandi, M. Marzioch, A. Bauer, J. Schultz, J. M. Rick, A. M. Michon, C. M. Cruciat, M. Remor, C. Hofert, M. Schelder, M. Brajenovic, H. Ruffner, A. Merino, K. Klein, M. Hudak, D. Dickson, T. Rudi, V. Gnau, A. Bauch, S. Bastuck, B. Huhse, C. Leutwein, M. A. Heurtier, R. R. Copley, A. Edelmann, E. Ouerfurth, V. Rybin, G. Drewes, M. Raida, T. Bouwmeester, P. Bork, B. Seraphin, B. Kuster, G. Neubauer, and G. Superti-Furga (2002) Functional organization of the yeast proteome by systematic analysis of protein complexes.Nature 415: 141–147.CrossRefGoogle Scholar
- Ho, Y., A. Gruhler, A. Heilbut, G. D. Bader, L. Moore, S. L. Adams, A. Millar, P. Taylor, K. Bennett, K. Boutilier, L. Y. Yang, C. Wolting, I. Donaldson, S. Schandorff, J. Shewnarane, M. Vo, J. Taggart, M. Goudreault, B. Muskat, C. Alfarano, D. Dewar, Z. Lin, K. Michalickova, A. R. Willems, H. Sassi, P. A. Nielsen, K. J. Rasmussen, J. R. Andersen, L. E. Johansen, L. H. Hansen, H. Jespersen, A. Podtelejnikov, E. Nielsen, J. Crawford, V. Poulsen, B. D. Sorensen, J. Matthiesen, R. C. Hendrickson, F. Gleeson, T. Pawson, M. F. Moran, D. Durocher, M. Mann, C. W. V. Hogue, D. Figeys, M. Tyers (2002) Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry.Nature 415: 180–183.CrossRefGoogle Scholar
- Raymackers, J., A. Daniels, V. De Brabandere, C. Missiaen, M. Dauwe, P. Verhaert, E. Vanmechelen, and L. Meheus (2000) Identification of two-dimensionally separated human cerebrospinal fluid proteins by N-terminal sequencing, matrix-assisted laser desorption/ionizationmass spectrometry, nanoliquid chromatography-electrospray ionization-time of flight-mass spectrometry, and tandem mass spectrometry.Electrophoresis 21: 2266–2283.CrossRefGoogle Scholar
- Wang, C., R. Oleschuk, F. Ouchen, J. Li, P. Thibault, and D. J. Harrison (2000) Integration of immobilized trypsin bead beds for protein digestion within a microfluidic chip incorporating capillary electrophoresis separations and an electrospray mass spectrometry interface.Rapid Commun. Mass Spectrom. 14: 1377–1383.CrossRefGoogle Scholar
- Viklund, C., E. Pontén, B. Glad, K. Irgum, P. Hörstedt, and F. Svec (1997) “Molded” macroporous poly (glycidyl methacrylate-co-trimethylolpropane trimethacrylate) materials with fine controlled porous properties: Preparation of monoliths using photoinitiated polymerization.Chem. Mater. 9: 463–471.CrossRefGoogle Scholar