A new approach for determination of neuropathy target esterase activity.
Neuropathy target esterase (NTE) was shown to be an excellent biochemical marker for screening of organophosphates (OPs) with respect to their ability to result in organophosphate induced delayed neurotoxicity (OPIDN). This paper describes a new biosensor approach to the analysis of NTE and its inhibitors. The method is based on the combination of NTE enzymatic hydrolysis of phenyl valerate (PV) with phenol detection by the Clark-type oxygen electrode modified by immobilized tyrosinase. The validity of this biosensor method is confirmed by the facts that the calibration curves for NTE obtained by colorimetric and flow-through electrochemical methods were nearly identical and the titration of NTE by test inhibitor mipafox was shown to yield the same pI50 values. The developed electrochemical methods can be considered as a promising approach both for serial express NTE analysis and for kinetic characteristics of NTE.
Sigolaeva
L V
LV
Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Russia. sens1rcmd@glasnet.ru
Eremenko
A V
AV
Makower
A
A
Makhaeva
G F
GF
Malygin
V V
VV
Kurochkin
I N
IN
D000818:Animals; D015374:Biosensing Techniques; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002645:Chickens; D004563:Electrochemistry / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004800:Enzymes, Immobilized; D005260:Female; D006868:Hydrolysis; D010755:Organophosphates / Q000633:toxicity; D010101:Oxygen Consumption; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014631:Valerates / Q000378:metabolism
D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D010755:Organophosphates; D014631:Valerates; C049867:phenyl valerate; D002265:Carboxylic Ester Hydrolases; C021859:neurotoxic esterase