Integrated microchip-device for the digestion, separation and postcolumn labeling of proteins and peptides.

Abstract

A microchip device was demonstrated that integrated enzymatic reactions, electrophoretic separation of the reactants from the products and post-separation labeling of proteins and peptides prior to detection. A tryptic digestion of oxidized insulin B-chain was performed in 15 min under stopped flow conditions in a heated channel, and the separation was completed in 1 min. Localized thermal control of the reaction channel was achieved using a resistive heating element. The separated reaction products were then labeled with naphthalene-2,3-dicarboxaldehyde (NDA) and detected by laser-induced fluorescence. A second reaction at elevated temperatures was also demonstrated for the on-chip reduction of disulfide bridges using insulin as a model protein. This device represents one of the highest levels, to date, of monolithic integration of chemical processes on a microchip.

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@article{Gottschlich2000IntegratedMF, title={Integrated microchip-device for the digestion, separation and postcolumn labeling of proteins and peptides.}, author={N Gottschlich and Christopher T Culbertson and Timothy E. Mcknight and Stephen C. Jacobson and John Michael Ramsey}, journal={Journal of chromatography. B, Biomedical sciences and applications}, year={2000}, volume={745 1}, pages={243-9} }