Integrated microchip-device for the digestion, separation and postcolumn labeling of proteins and peptides.
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.
Gottschlich
N
N
Oak Ridge National Laboratory, TN 37831-6142, USA.
Culbertson
C T
CT
McKnight
T E
TE
Jacobson
S C
SC
Ramsey
J M
JM
D006868:Hydrolysis; D008904:Miniaturization; D009281:Naphthalenes / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012666:Semiconductors
D009281:Naphthalenes; D010455:Peptides; D011506:Proteins; C027717:2,3-naphthalenedicarboxaldehyde