PMID: 11906264Mar 22, 2002Paper

Cysteine-free mutant of aequorin as a photolabel in immunoassay development

Bioconjugate Chemistry
Suresh ShresthaSylvia Daunert

Abstract

The bioluminescent protein aequorin is a sensitive label that has been employed in a number of analytical applications. A mutant of aequorin with enhanced stability produced recombinantly in our laboratory has been employed as a label in the development of an immunoassay for digoxin. Digoxin is a cardiac glycoside used in the treatment of congestive heart failure. This drug has a very narrow therapeutic range of 0.8-2.0 ng/mL (1.0-2.5 nmol/L), thus requiring therapeutic drug monitoring. In this study, a derivative of digoxigenin was chemically conjugated to the mutant aequorin, and the resulting protein-digoxigenin derivative conjugates were characterized in terms of their luminescence properties. A solid-phase immunoassay for digoxin was then developed. The detection limit of the assay for digoxin was 1 x 10(-12) M. To demonstrate the use of this mutant aequorin as a label in biological sample analysis without any need for pretreatment of the samples, the assay was tested in serum spiked with digoxin. Interference from digoxin analogues was also evaluated to determine the specificity of the assay.

References

Feb 15, 1991·The American Journal of Cardiology·M S ChernD L Wu
Jan 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·K KuroseF I Tsuji
Apr 1, 1997·Clinical Pharmacology and Therapeutics·M C TzouR A Sams
Dec 24, 1997·Analytical Chemistry·C L CrofcheckS Daunert
Jun 11, 1998·Trends in Biotechnology·J M Kendall, M N Badminton
Mar 2, 1999·Therapeutic Drug Monitoring·B K DeW L Roberts
Nov 11, 1999·Trends in Biotechnology·K JonesM Keenan
Jun 17, 2000·Analytical Chemistry·A L GrosvenorK W Anderson
Nov 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·F I TsujiY Sakaki

❮ Previous
Next ❯

Citations

Jun 2, 2018·Photochemistry and Photobiology·Elena V Eremeeva, Eugene S Vysotski
Aug 3, 2014·Analytical and Bioanalytical Chemistry·Xiaoge QuSylvia Daunert
Dec 10, 2008·Bioconjugate Chemistry·Kyle A CissellSapna K Deo
Sep 18, 2003·Bioconjugate Chemistry·Kyriaki GlynouTheodore K Christopoulos
Feb 27, 2018·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Sana SharifianKhosro Khajeh

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cardiac Glycosides

Cardiac glycosides are a diverse family of naturally derived compounds that bind to and inhibit na+/k+-atpase. Discover the latest research on cardiac glycosides heres.