Application of a long-term enhanced xanthine oxidase-induced luminescence in solid-phase immunoassays

Analytical Biochemistry
A BaretJ Aumaille

Abstract

The use of xanthine oxidase (XO) as a label in immunoanalysis has not been previously reported. This can be explained by the difficulties encountered in XO assays (poor sensitivity and versatility) and the competitive inhibition of the enzyme by allopurinol, a widely used hypouricemic agent. We demonstrate here that both difficulties can be circumvented. (i) The XO-dependent luminescent signal related to the oxidation of luminol is dramatically enhanced in the presence of iron-EDTA complex and sodium perborate in alkaline buffer. The mechanism of this enhancement is consistent with an O2-driven Fenton reaction, leading to the production of highly reactive OH radical. (ii) Residual inhibition of solid-phase bound XO by serum allopurinol and its metabolites is spontaneously reversible and can be prevented by the presence of folic acid or azahypoxanthine in the incubation buffer. With these two problems solved, XO can be classified as a choice label in immunoanalysis with the following properties: (i) high detection sensitivity (3 amol label), (ii) long-term luminescent signal (several days), (iii) versatile preparation and stability of conjugates, and (iv) long-term stability of the luminescence reagent. As an example of applicat...Continue Reading

References

Jan 1, 1989·Free Radical Biology & Medicine·H C Sutton, C C Winterbourn
May 1, 1986·Archives of Biochemistry and Biophysics·B Halliwell, J M Gutteridge
Jan 1, 1987·Free Radical Biology & Medicine·H B Dunford
Jun 1, 1986·Journal of Bioluminescence and Chemiluminescence·R HaggartT P Whitehead
Sep 1, 1986·Clinical Pharmacokinetics·G A Murrell, W G Rapeport
Jan 1, 1985·Molecular Aspects of Medicine·B Halliwell, J M Gutteridge
Dec 1, 1973·Photochemistry and Photobiology·E K Hodgson, I Fridovich

❮ Previous
Next ❯

Citations

Jul 1, 1995·Journal of Bioluminescence and Chemiluminescence·L LamarcqE Brambilla
Jun 24, 1992·Journal of Immunological Methods·Y ChikhaouiJ C Nicolas
Oct 16, 1996·Journal of Immunological Methods·H A RongenW P van Bennekom
Jun 30, 1994·Journal of Biotechnology·C Kessler
Apr 1, 1994·Journal of Pharmaceutical and Biomedical Analysis·H A RongenW P van Bennekom
Apr 1, 1991·Current Opinion in Immunology·P Tijssen, A Adam
Feb 1, 1991·Current Opinion in Biotechnology·T T Ngo
Oct 31, 2008·Talanta·C DodeigneR Lejeune
Apr 26, 2011·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Bogdan Alexandru StoicaMihai Nechifor
Apr 4, 2018·Cold Spring Harbor Protocols·Larry J Kricka

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.