Abstract
Histamine is a biogenic amine synthesized by the enzymatic decarboxylation of histidine. Implication of histamine in allergy is well described but histamine is also found in some specific neurones, functions as a neurotransmitter and regulates sleep/wake cycles, hormonal secretion, cardiovascular control and thermo-regulation. We have developed a TR-FRET histamine assay, based on the competition between sample histamine and allophycocyanine (XL665) labelled histamine for binding to a Europium cryptate (EuK) labelled antibody. As histamine is a small monoamine molecule, high affinity antibodies have been raised against carrier protein conjugated histamine. Therefore, sample histamine needs to be derivatized in the same way as the conjugated histamine, so that the antibody will have a similar affinity for both molecules. This acylation step is performed directly in wells and does not need to be done in separate vials, making handling easier for large numbers of samples. The incubation takes place at room temperature for 3 hours. The assay covers a measurement range of 1.56 to 400 nM and shows an analytical sensitivity of 1.3nM. We have shown that miniaturization of sample and reagents volumes down to 20μl does not alter these performances. This histamine release assay provides a particularly well adapted procedure for HTS and secondary screening compared to current heterogeneous methods.
Keywords: Homogeneous assay, homogeneous time resolved fluorescence, europium cryptate, histamine
Combinatorial Chemistry & High Throughput Screening
Title: Homogeneous Time Resolved Fluorescence Assay to Measure Histamine Release
Volume: 6 Issue: 8
Author(s): Emmanuel J. Claret, Josy Ouled-Diaf and Patrick Seguin
Affiliation:
Keywords: Homogeneous assay, homogeneous time resolved fluorescence, europium cryptate, histamine
Abstract: Histamine is a biogenic amine synthesized by the enzymatic decarboxylation of histidine. Implication of histamine in allergy is well described but histamine is also found in some specific neurones, functions as a neurotransmitter and regulates sleep/wake cycles, hormonal secretion, cardiovascular control and thermo-regulation. We have developed a TR-FRET histamine assay, based on the competition between sample histamine and allophycocyanine (XL665) labelled histamine for binding to a Europium cryptate (EuK) labelled antibody. As histamine is a small monoamine molecule, high affinity antibodies have been raised against carrier protein conjugated histamine. Therefore, sample histamine needs to be derivatized in the same way as the conjugated histamine, so that the antibody will have a similar affinity for both molecules. This acylation step is performed directly in wells and does not need to be done in separate vials, making handling easier for large numbers of samples. The incubation takes place at room temperature for 3 hours. The assay covers a measurement range of 1.56 to 400 nM and shows an analytical sensitivity of 1.3nM. We have shown that miniaturization of sample and reagents volumes down to 20μl does not alter these performances. This histamine release assay provides a particularly well adapted procedure for HTS and secondary screening compared to current heterogeneous methods.
Export Options
About this article
Cite this article as:
Claret J. Emmanuel, Ouled-Diaf Josy and Seguin Patrick, Homogeneous Time Resolved Fluorescence Assay to Measure Histamine Release, Combinatorial Chemistry & High Throughput Screening 2003; 6 (8) . https://dx.doi.org/10.2174/138620703771826892
DOI https://dx.doi.org/10.2174/138620703771826892 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
Integrating Network Pharmacology and Traditional Medicine: A New Perspective in Drug Mechanism Research
Network pharmacology is a network construction and network topology analysis strategy that combines pharmacology and pharmacodynamics. In recent years, network pharmacology has emerged as a powerful tool that can be integrated with pharmacology. Natural products commonly function in multicomponent, multitarget, and multipathway systems. Some examples encompass Ayurveda, traditional Chinese medicines ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Acute Phase Proteins In Acute Coronary Syndrome: An up-to-date
Cardiovascular & Hematological Agents in Medicinal Chemistry Betaine Modulating MIF-Mediated Oxidative Stress, Inflammation and Fibrogenesis in Thioacetamide-Induced Nephrotoxicity
Current Medicinal Chemistry Association of Oxidative Stress to the Genesis of Anxiety: Implications for Possible Therapeutic Interventions
Current Neuropharmacology Genistein Aglycone: A Dual Mode of Action Anti-Osteoporotic Soy Isoflavone Rebalancing Bone Turnover Towards Bone Formation
Current Medicinal Chemistry Lycopene Modulation of Molecular Targets Affected by Smoking Exposure
Current Cancer Drug Targets The Endocannabinoid Signaling System: A Potential Target for Next- Generation Therapeutics for Alcoholism
Mini-Reviews in Medicinal Chemistry Medical Treatment of Chronic Venous Insufficiency
Vascular Disease Prevention (Discontinued) Editorial (Advances and Controversies in Stem Cell Therapies and Tissue Engineering Strategies Applicable to Trauma and Orthopaedic Surgery)
Current Stem Cell Research & Therapy Haploinsufficiency of DNA Damage Response Genes and their Potential Influence in Human Genomic Disorders
Current Genomics Endothelial Dysfunction, Obesity and Insulin Resistance
Current Vascular Pharmacology Histone Deacetylase Inhibitors: Molecular and Biological Activity as a Premise to Clinical Application
Current Drug Metabolism Novel Carriers for Controlled Site Specific Delivery of Anti-Inflammatory Agents
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Medicinal Chemistry and Anti-Inflammatory Activity of Nitric Oxide-Releasing NSAI Drugs
Mini-Reviews in Medicinal Chemistry Neuroprotective Effects of Exercise Treatments After Injury: The Dual Role of Neurotrophic Factors
Current Neuropharmacology Tissue Transglutaminase Expression During Neural Differentiation of Human Mesenchymal Stem Cells
CNS & Neurological Disorders - Drug Targets The Potential Role of Erythropoietin as a Pleiotropic Agent in Post-cardiac Arrest Syndrome
Current Pharmaceutical Design Anti-carcinogenic Effect of Cathepsin K Inhibitor, Odanacatib with a Low Dose of Cisplatin Against Human Breast Carcinoma MCF-7 and MDA-MB231 Cells
Current Cancer Therapy Reviews Do We Need More Research on Platelet Function in South Asian Immigrants?
Inflammation & Allergy - Drug Targets (Discontinued) A New Twist in Cellular Resistance to the Anticancer Drug Bleomycin-A5
Current Drug Metabolism Pharmaceutical Approaches for Treatment of Hepatitis C virus
Current Pharmaceutical Design