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Quantitation of solid-phase immunoassays using a fibre-optic reflectance spectrophotometer
D McCarthy1, A Rehman, D N Batchelder
1School of Biological Sciences, Queen Mary and Westfield College, London, U.K.
Journal of Immunological Methods
|October 25, 1991
Summary
A new fiber-optic spectrometer accurately quantifies solid-phase immunoassays by measuring colored products. This method provides results comparable to established techniques for detecting rheumatoid factor in serum.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Immunology
Background:
- Immunoassays often produce colored products on solid phases.
- Quantifying these colored products is crucial for accurate diagnostic results.
- Existing methods for quantitation may have limitations.
Purpose of the Study:
- To describe a novel reflectance measuring instrument for quantitating solid-phase immunoassays.
- To present reflectance spectra of common chromophores used in these assays.
- To evaluate the instrument's performance against established methods.
Main Methods:
- Development of a fiber-optic spectrometer linked to a personal computer.
- Measurement of reflectance spectra of chromophores on solid phases.
- Application in silver-enhanced solid-phase immunogold capture assays for IgG and IgM rheumatoid factor (RF).
- Comparison with rate nephelometry and latex agglutination assays.
Main Results:
- The instrument successfully quantitates solid-phase immunoassays producing colored chromophores.
- Reflectance spectra of common assay chromophores were characterized.
- Results for IgG and IgM RF determination were comparable to established methods.
Conclusions:
- Fiber-optic reflectance photometry is an accurate and useful method for quantitating solid-phase immunoassays.
- This technique offers a reliable alternative to existing diagnostic procedures.
- The developed instrument demonstrates significant potential for clinical diagnostics.