A new way in homogeneous immunoassay: reversed micellar systems as a medium for analysis.

Analytical Biochemistry
A V KabanovA V Levashov

Abstract

Possibilities of a new principle for the homogeneous enzyme immunoassay utilizing the systems of surfactant reversed micelles in organic solvents have been demonstrated taking thyroxine determination as an example. The catalytic activity of an enzyme, solubilized in such systems, is determined by the ratio of geometric dimensions of the micellar matrice and the enzyme molecule. The addition of antibodies against thyroxine to the peroxidase-thyroxine conjugate, solubilized in the system of reversed micelles of aerosol OT in octane, leads to the formation of the immune complex whose size differs substantially from that of the initial enzyme-antigen conjugate. This induces changes in the peroxidase catalytic activity. The addition of free thyroxine to the system stimulates the conjugate release from the immune complex and, consequently, the reduction of the peroxidase activity to the initial level. Sensitivity of the analysis in reversed micellar systems can be regulated by changing the surfactant hydration degree. Substances of different nature (both hydrophobic and hydrophylic) can be solubilized in reverse micellar systems under standard conditions, which allows determination of water insoluble antigens.

References

Mar 17, 1986·European Journal of Biochemistry·K MartinekI V Berezin
Jun 6, 1989·Biochimica Et Biophysica Acta·K MartinekA V Levashov
Feb 24, 1988·Biochimica Et Biophysica Acta·P L LuisiB H Robinson
Sep 1, 1971·The Biochemical Journal·R Fields

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Citations

Nov 1, 1990·Applied Microbiology and Biotechnology·B MirhabibollahiR G Kroll
Sep 9, 1996·Journal of Immunological Methods·K NielsenF Thomas
Feb 1, 1991·Current Opinion in Biotechnology·T T Ngo
Aug 31, 1992·Biochemical and Biophysical Research Communications·R M Uppu, W A Pryor
Mar 31, 1995·Annals of the New York Academy of Sciences·A V Levashov, N Klyachko

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