Enzyme kinetics by directly imaging a porous silicon microfluidic reactor using desorption/ionization on silicon mass spectrometry.
Enzyme kinetics were obtained in a porous silicon microfluidic channel by combining an enzyme and substrate droplet, allowing them to react and deposit a small amount of residue on the channel walls, and then analyzing this residue by directly ionizing the channel walls using a matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) laser source. The porous silicon of the channel walls functions in a manner analogous to the matrix in MALDI-MS, and is referred to as a desorption/ionization on silicon mass spectrometry (DIOS-MS) target when used in this configuration. Mass spectrometry signal intensity of substrate residue correlates with relative concentration, and position in the microchannel correlates with time, thus allowing determination of kinetic parameters. The system is especially suitable for initial reaction velocity determination. This microreactor is broadly applicable to time-resolved kinetic assays as long as at least one substrate or product of the reaction is ionizable by DIOS-MS.
MESA+ Institute for Nanotechnology, Mesoscale Chemical Systems, Meander 151, University of Twente, Postbus 217, 7500 AE Enschede, The Netherlands.
Kevin P
KP
Nichols
Seyla
S
Azoz
Han J G E
HJ
Gardeniers
D000818:Animals; D001119:Arginase / Q000032:analysis* / Q000378:metabolism*; D002417:Cattle; D007477:Ions / Q000737:chemistry; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D016062:Porosity; D012825:Silicon / Q000737:chemistry*
D007477:Ions; D001119:Arginase; D012825:Silicon