The measurement of alkaline phosphatase at nanomolar concentration within 70 s using a disposable microelectrochemical transistor

Bioelectrochemistry. 2004 Aug;64(1):53-9. doi: 10.1016/j.bioelechem.2004.01.004.

Abstract

We report a new approach to the measurement of alkaline phosphatase concentration based on the use of a disposable poly(aniline) microelectrochemical transistor. The measurement is carried out in a two cell configuration in which the poly(aniline) microelectrochemical transistor operates in acid solution and is connected to the alkaline buffer solution containing the analyte by a salt bridge. Disposable microelectrochemical transistors were reproducibly fabricated by electrochemical deposition of poly(aniline) onto photolithographically fabricated gold microband arrays. Using these devices alkaline phosphatase was detected by employing p-aminophenyl phosphate as the substrate for the enzyme and using glucose and glucose oxidase to recycle the p-aminophenol generated upon enzyme catalysed hydrolysis of the phosphate. Recycling the p-aminophenol with glucose and glucose oxidase amplified the detection of alkaline phosphatase approximately tenfold. Using this approach we obtain linear calibration curves for alkaline phosphatase up to 5 nM within 70 s on single use devices.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / analysis*
  • Aminophenols / metabolism
  • Aniline Compounds / chemistry
  • Calibration
  • Catalysis
  • Electrochemistry / methods*
  • Glucose / analysis
  • Glucose Oxidase / analysis
  • Gold / chemistry
  • Hydrolysis
  • Microchemistry
  • Polymers / chemistry
  • Sensitivity and Specificity
  • Time Factors
  • Transistors, Electronic*

Substances

  • Aminophenols
  • Aniline Compounds
  • Polymers
  • polyaniline
  • Gold
  • Glucose Oxidase
  • Alkaline Phosphatase
  • Glucose
  • 4-aminophenol