A two site electrochemical enzyme immunoassay for thyrotropin (TSH) has been developed. This assay is based on the use of an immobilized capture antibody and a biotinylated second antibody. Detection is achieved via avidin labelled with alkaline phosphatase. The substrate 1-naphthyl phosphate was used and the product 1-naphthol was detected at disposable screen-printed carbon 8-electrode combs using specially designed instrumentation.
ThompsonRQPorterMStuverCHalsallHBHeinemanWRBuckleyEZeptomole detection limit for alkaline phosphatase using 4-amino-phenylphosphate, amperometric detection, and an optimal buffer system. Anal Chim Acta1993; 271: 223–9
2.
PriceCPNewmanDJPrinciples and Practice of Immunoassay, 1st edn.New York: Stockton Press, 1991
3.
DoyleMJHalsallHBHeinemanWREnzyme-linked immunoadsorbent assay with electrochemical detection for an α1-acid glycoprotein. Anal Chem1984; 56: 2355–60
4.
WehmeyerKRHalsallHBHeinemanWRHeterogeneous enzyme immunoassay with electrochemical detection; competitive and ‘sandwich’ type immunoassays. Clin Chem1985; 31: 1546–9
5.
TangHLunteCHalsallHBHeinemanWRp-Aminophenyl phosphate: an improved substrate for electrochemical enzyme immunoassay. Anal Chim Acta1988; 244: 187–94
6.
CardosiMBirchSTalbotJPhillipsAAn electrochemical immunoassay for Clostridium perfringens phospholipase C. Electroanalysis1991; 3: 169–76
7.
McNeilCJHigginsIJBannisterJVAmperometric determination of alkaline phosphatase activity: Application to enzyme immunoassay. Biosensors1988; 3: 199–209
8.
MossDWA note on the spectrophotometric estimation of alkaline phosphatase activity. Enzymologia1966; 31: 193–202
9.
McNeilCJBannisterJVAtheyDMethod and apparatus for electrochemical immunoassay. European Patent Application1990; 89312157.4
10.
GodingJWMonoclonal Antibodies: Principles and Practice. New York: Wiley, 1989
11.
AtheyDPhD Thesis1993, University of Newcastle Upon Tyne
12.
TieFPanARuBWangWHuYAn improved ELISA with linear sweep voltammetry detection. J Immunol Methods1992; 149: 115–20
13.
BlandJMAltmanDGStatistical methods for assessing agreement between two methods of clinical measurement. Lancet1986; i: 307–10
14.
ViljaPFlexibility of non-competitive avidin-biotin immunoassay: immunoassay of lutropin applied to different signal detection systems. Clin Chem1990; 36: 1897–901
15.
FarinaLIacobelloCOrlandiniSIusAAlbertiniAThe effect of the concentration ratio of avidin and biotin on a single step sandwich immunoassay. Ann Clin Biochem1993; 30: 87–9
