title: 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.
authors.affiliation: Oak Ridge National Laboratory, TN 37831-6142, USA.
authors.forename: N
authors.initials: N
authors.lastname: Gottschlich
authors.forename: C T
authors.initials: CT
authors.lastname: Culbertson
authors.forename: T E
authors.initials: TE
authors.lastname: McKnight
authors.forename: S C
authors.initials: SC
authors.lastname: Jacobson
authors.forename: J M
authors.initials: JM
authors.lastname: Ramsey
mesh_terms: D006868:Hydrolysis; D008904:Miniaturization; D009281:Naphthalenes / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012666:Semiconductors
chemical_list: D009281:Naphthalenes; D010455:Peptides; D011506:Proteins; C027717:2,3-naphthalenedicarboxaldehyde