Dual-enzyme cascade--an amplified method for the detection of alkaline phosphatase.
A method in which a two-enzyme cascade is used for rapid and sensitive detection of alkaline phosphatase is described. A masked inhibitor, 4-(3-oxo-4,4,4-trifluorobutyl)phenyl phosphate, is dephosphorylated by the action of alkaline phosphatase. The resulting compound, 1,1,1-trifluoro-4-(4-hydroxyphenyl)-butan-2-one, acts as a potent inhibitor of the second enzyme, a liver carboxylesterase. A determination of the residual esterase activity provides a highly sensitive indication of the original phosphatase concentration. The sensitivity of this dual-enzyme cascade is approximately 125-fold greater than that observed for the direct detection of phosphatase activity with p-nitrophenyl phosphate.
Becton Dickinson Research Center, Research Triangle Park, North Carolina 27709-2016.
P D
PD
Mize
R A
RA
Hoke
C P
CP
Linn
J E
JE
Reardon
T H
TH
Schulte
D000469:Alkaline Phosphatase / Q000032:analysis* / Q000037:antagonists & inhibitors; D055598:Chemical Phenomena; D002621:Chemistry; D004950:Esterases / Q000378:metabolism
D004950:Esterases; D000469:Alkaline Phosphatase