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Amperometric enzyme-amplified immunoassays.

C J Stanley¹, R B Cox, M F Cardosi

¹Novo Biolabs Ltd., Downham House, Cambridge, U.K.

Journal of Immunological Methods

|September 13, 1988

Summary

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Electrochemical enzyme-amplified immunoassays offer enhanced dynamic range compared to optical methods. These novel amperometric techniques show promise for replacing traditional optical immunoassays in diagnostics.

Area of Science:

Biomedical Engineering
Analytical Chemistry
Electrochemistry

Background:

Enzyme-amplified immunoassays are widely used for sensitive detection.
Electrochemical detection offers potential advantages in sensitivity and dynamic range.
Adapting immunoassays for electrochemical measurement requires efficient signal transduction.

Purpose of the Study:

To adapt enzyme-amplified immunoassays for electrochemical measurement.
To evaluate amperometric methods using different electrode systems.
To compare the performance of electrochemical immunoassays with optical techniques.

Main Methods:

Utilized an NAD+/NADH redox cycle coupled to an electrode via diaphorase.
Employed two amperometric methods: an organic conducting salt electrode (NMP+/TCNQ-) and a platinum wire with ferricyanide mediator.

Related Experiment Videos

Applied the methods to an immunoenzymometric assay for human prostatic acid phosphatase.

Main Results:

Achieved sensitivities comparable to existing optical techniques for human prostatic acid phosphatase detection.
Demonstrated an increased dynamic range of at least two orders of magnitude for the electrochemical assays.
Successfully coupled the NAD+/NADH redox cycle to electrodes via diaphorase for signal amplification.

Conclusions:

Electrochemical enzyme-amplified immunoassays provide a viable alternative to optical methods.
The developed amperometric techniques offer significantly improved dynamic range.
Electrochemical immunoassays hold potential for future diagnostic applications, possibly replacing optical counterparts.