PMID: 7017002Jan 1, 1981Paper

Homogeneous substrate-labeled fluorescent immunoassay for IgG in human serum

Journal of Immunological Methods
T T NgoJ F Burd

Abstract

A homogeneous substrate-labeled fluorescent immunoassay for IgG has been developed. Purified IgG was covalently labeled with 6-(7-beta-galactosylcoumarin-3-carboxamido)-hexylamine to form a stable conjugate, GU-IgG. The galactosyl residue was hydrolyzed from GU-IgG by beta-galactosidase and the progress of the hydrolysis was monitored by the increase in fluorescence emission at 450 nm with excitation at 400 nm. Antibody to IgG diminished the activity of GU-IgG as a substrate for beta-galactosidase. Competitive binding immunoassays were conducted by allowing added IgG and GU-IgG to compete for a limited number of antibody binding sites. Hence, the fluorescence produced by enzymic hydrolysis increased with the level of added IgG. This method provides a simple and reliable immunoassay for IgG and is applicable to other proteins.

References

May 26, 1972·Biochemical and Biophysical Research Communications·K E RubensteinE F Ullman
Feb 1, 1980·Analytical Biochemistry·I GibbonsE F Ullman

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Citations

Apr 16, 1982·Molecular and Cellular Biochemistry·T T Ngo, H M Lenhoff
Sep 1, 1982·Applied Biochemistry and Biotechnology·R C Boguslaski, T M Li
Dec 15, 1989·Clinica Chimica Acta; International Journal of Clinical Chemistry·P L KhannaJ D Harris
Jan 1, 1983·The International Journal of Biochemistry·T T Ngo
Jun 24, 1992·Journal of Immunological Methods·T Porstmann, S T Kiessig
Jun 24, 1992·Journal of Immunological Methods·S H Jenkins
Jan 1, 1983·Journal of Immunoassay·J PatinkinB Fridlender
Aug 12, 1983·Biochemical and Biophysical Research Communications·T T Ngo, H M Lenhoff
Feb 1, 1984·Analytical Biochemistry·C Bacquet, D Y Twumasi

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