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Proceedings Paper

Enzymatic digestion and liquid chromatography in micropillar reactors: hydrodynamic versus electro-osmotic flow
Author(s): Nicolas Sarrut; Stephanie Bouffet; Frederique Mittler; Olivier Constantin; P. Combette; Jan Sudor; Florence Ricoul; Francoise Vinet; Jerome Garin; Claude Vauchier
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Paper Abstract

Enzymatic digestion and peptide separation are basic steps for preparation of protein samples prior to their analysis by mass spectrometry. Micro-pillar reactors for digestion and reverse phase liquid chromatography were designed and constructed using semiconductor technologies. The performances of the micro-machined reactors were evaluated: complete Cytochrome C digestion was achieved in 6 min for a concentration up to 25 pmol.μl-1 and the separation micro-column was seen to exhibit separation capabilities and capacity close to those obtained with a commercial column. Furthermore, a comparative study between hydrodynamic and electroosmotic driven flows was performed for each peptide of a Cytochrome C digest. It was demonstrated that parasitic electrophoretic phenomena disturbed peptide mobility but not protein identification.

Paper Details

Date Published: 22 January 2005
PDF: 11 pages
Proc. SPIE 5718, Microfluidics, BioMEMS, and Medical Microsystems III, (22 January 2005); doi: 10.1117/12.590150
Show Author Affiliations
Nicolas Sarrut, CEA-LETI Grenoble (France)
Stephanie Bouffet, CEA Grenoble (France)
Frederique Mittler, CEA-LETI Grenoble (France)
Olivier Constantin, CEA-LETI Grenoble (France)
P. Combette, CEA Grenoble (France)
Jan Sudor, CEA-LETI Grenoble (France)
Florence Ricoul, CEA-LETI Grenoble (France)
Francoise Vinet, CEA-LETI Grenoble (France)
Jerome Garin, CEA Grenoble (France)
Claude Vauchier, CEA-LETI Grenoble (France)

Published in SPIE Proceedings Vol. 5718:
Microfluidics, BioMEMS, and Medical Microsystems III
Ian Papautsky; Isabelle Chartier, Editor(s)

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