ArticleBiophysics (Nagoya-shi, Japan)2014
Ultrasensitive enzyme-linked immunosorbent assay (ELISA) of proteins by combination with the thio-NAD cycling method.
Article in Biophysics (Nagoya-shi, Japan), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 43 citations in OpenAlex.
- Next-Generation Diagnostic Technologies for Dengue Virus Detection: Microfluidics, Biosensing, CRISPR, and AI Approaches.Sensors (Basel, Switzerland) · 2025Review
- Ultrasensitive ELISA for Accurate Detection ofOpen forum infectious diseases · 2025Article
- An overview of ELISA: a review and update on best laboratory practices for quantifying peptides and proteins in biological fluids.The Journal of international medical research · 2025Review
- Quantification of HPV16 E7 Oncoproteins in Urine Specimens from Women with Cervical Intraepithelial Neoplasia.Microorganisms · 2024Article
- Ultrasensitive protein-level detection for respiratory infectious viruses.Frontiers in immunology · 2024Article
- Advanced Detection Method for Dengue NS1 Protein Using Ultrasensitive ELISA with Thio-NAD Cycling.Viruses · 2023Article
- Protein Quantitation and Analysis of Purity.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Gastric Cancer Cell-Derived Exosomal GRP78 Enhances Angiogenesis upon Stimulation of Vascular Endothelial Cells.Current issues in molecular biology · 2022Article
- Article
- Improved Conventional and New Approaches in the Diagnosis of Tuberculosis.Frontiers in microbiology · 2022Review
- Modified ELISA for Ultrasensitive Diagnosis.Journal of clinical medicine · 2021Review
- Ultrasensitive Detection of SARS-CoV-2 Spike Proteins Using the Thio-NAD Cycling Reaction: A Preliminary Study before Clinical Trials.Microorganisms · 2021Article
- Zeptomole Detection of an Enzyme by a Simple Colorimetric Method.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2021Article
- Antigen tests for COVID-19.Biophysics and physicobiology · 2021Article
- A novel, rapid (within hours) culture-free diagnostic method for detecting live Mycobacterium tuberculosis with high sensitivity.EBioMedicine · 2020Article
- Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2.Diagnostics (Basel, Switzerland) · 2020Article
- Ultrasensitive ELISA Developed for Diagnosis.Diagnostics (Basel, Switzerland) · 2019Review
- A novel role of oxytocin: Oxytocin-induced well-being in humans.Biophysics and physicobiology · 2019Article
- Urinary adiponectin as a new diagnostic index for chronic kidney disease due to diabetic nephropathy.BMJ open diabetes research & care · 2019Article
- Function of insulin in snail brain in associative learning.Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology · 2015Review
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
An ultrasensitive method for the determination of proteins is described that combines an enzyme-linked immunosorbent assay (ELISA) and a thionicotinamide-adenine dinucleotide (thio-NAD) cycling method. A sandwich method using a primary and a secondary antibody for antigens is employed in an ELISA. An androsterone derivative, 3α-hydroxysteroid, is produced by the hydrolysis of 3α-hydroxysteroid 3-phosphate with alkaline phosphatase linked to the secondary antibody. This 3α-hydroxysteroid is oxidized to a 3-ketosteroid by 3α- hydroxysteroid dehydrogenase (3α-HSD) with a cofactor thio-NAD. By the opposite reaction, the 3-ketosteroid is reduced to a 3α-hydroxysteroid by 3α-HSD with a cofactor NADH. During this cycling reaction, thio-NADH accumulates in a quadratic function-like fashion. Accumulated thio-NADH can be measured directly at an absorbance of 400 nm without any interference from other cofactors. These features enable us to detect a target protein with ultrasensitivity (10(-19) mol/assay) by measuring the cumulative quantity of thio-NADH. Our ultrasensitive determination of proteins thus allows for the detection of small amounts of proteins only by the application of thio-NAD cycling reagents to the usual ELISA system.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.