Document Type

Article

Publication Date

1989

Publication Title

Journal of Pharmaceutical and Biomedical Analysis

Abstract

A sandwich electrochemical enzyme immunoassay with flow injection analysis for the model antigen mouse IgG has been developed with alkaline phosphatase as the enzyme label. The enzyme substrate, 4-aminophenyl phosphate and its enzymatic reaction product, 4-aminophenol have been studied by cyclic and hydrodynamic voltammetry. The determination of 4-aminophenol by flow injection analysis with electrochemical detection (FIAEC) has a linear range of 5.0 × 10−8 to 1.0 × 10−5 M, a detection limit of 2.4 × 10−8 M, and a sample throughput of 72 samples/h. The detection limit is set by a background capacitance response, which depends on the ionic strength difference between the sample and the mobile phase. The sandwich immunoassay has been characterized with respect to substrate concentration for the enzymatic reaction, detection limit, dynamic range and sources of error. Mouse IgG can be determined with a detection limit of 0.81 pg ml−1 by a 30-min substrate incubation time and a six orders of magnitude linear dynamic range.

Comments

This work was funded by NSF grant CHE 8217045.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

DOI

10.1016/0731-7085(89)80136-0

Version

Postprint

Issue

12

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