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A protein assay based on colloidal gold conjugates with trypsin.
Lev A Dykman1, Vladimir A Bogatyrev, Boris N Khlebtsov
1Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 13 Prospekt Entuziastov, Saratov 410049, Russia.
Analytical Biochemistry
|May 4, 2005
Summary
A new sol particle immunoassay (SPIA) uses microtitration plates and an enzyme-linked immunosorbent assay reader for protein detection. This novel method enhances gold nanoparticle conjugate analysis for immunoglobulin G and trypsin quantification.
Area of Science:
- Nanotechnology
- Biochemistry
- Analytical Chemistry
Background:
- The standard sol particle immunoassay (SPIA) relies on gold nanoparticle aggregation and spectrophotometry.
- Existing methods require specialized equipment for spectrophotometric analysis.
Purpose of the Study:
- To develop a novel SPIA format using microtitration plates and an enzyme-linked immunosorbent assay (ELISA) reader.
- To adapt SPIA for quantifying immunoglobulin G (IgG) and trypsin.
Main Methods:
- Utilized 15-nm colloidal gold-protein A conjugates for IgG determination.
- Developed a sol particle-trypsin assay with gold nanoparticle-trypsin conjugates.
- Measured spectral extinction changes upon protein addition to assess gold nanoparticle aggregation.
Main Results:
- Demonstrated the efficacy of the novel SPIA format for IgG quantification.
- Successfully applied the sol particle-trypsin assay for trypsin detection.
- Observed spectral extinction changes correlated with protein-induced gold nanoparticle aggregation.
Conclusions:
- The novel SPIA format offers a viable alternative to conventional spectrophotometry.
- This method provides a sensitive approach for protein detection using gold nanoparticle conjugates.
- The assay principle is based on protein-mediated aggregation of gold nanoparticles, detectable via spectral changes.