TY - JOUR
T1 - Enzymatic reactions in microfluidic devices
T2 - Michaelis-Menten kinetics
AU - Ristenpart, William D.
AU - Wan, Jiandi
AU - Stone, Howard A.
PY - 2008/5/1
Y1 - 2008/5/1
N2 - Kinetic rate constants for enzymatic reactions are typically measured with a series of experiments at different substrate concentrations in a well-mixed container. Here we demonstrate a microfluidic technique for measuring Michaelis-Menten rate constants with only a single experiment. Enzyme and substrate are brought together in a coflow microfluidic device, and we establish analytically and numerically that the initial concentration of product scales with the distance x along the channel as x5/2. Measurements of the initial rate of product formation, combined with the quasi-steady rate of product formation further downstream, yield the rate constants. We corroborate the x5/2 scaling result experimentally using the bioluminescent reaction between ATP and luciferase/luciferin as a model system.
AB - Kinetic rate constants for enzymatic reactions are typically measured with a series of experiments at different substrate concentrations in a well-mixed container. Here we demonstrate a microfluidic technique for measuring Michaelis-Menten rate constants with only a single experiment. Enzyme and substrate are brought together in a coflow microfluidic device, and we establish analytically and numerically that the initial concentration of product scales with the distance x along the channel as x5/2. Measurements of the initial rate of product formation, combined with the quasi-steady rate of product formation further downstream, yield the rate constants. We corroborate the x5/2 scaling result experimentally using the bioluminescent reaction between ATP and luciferase/luciferin as a model system.
UR - http://www.scopus.com/inward/record.url?scp=42949124472&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=42949124472&partnerID=8YFLogxK
U2 - 10.1021/ac702469u
DO - 10.1021/ac702469u
M3 - Article
C2 - 18355085
AN - SCOPUS:42949124472
SN - 0003-2700
VL - 80
SP - 3270
EP - 3276
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 9
ER -