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| Enzyme-linked immunosorbent assay | |
|---|---|
| Name | Enzyme-linked immunosorbent assay |
| Caption | Microplate wells used in immunoassay |
| Invented | 1971 |
| Inventor | Peter Perlmann; Eva Engvall |
| Field | Immunology |
Enzyme-linked immunosorbent assay is a laboratory technique for detecting and quantifying antigens or antibodies using enzyme-conjugated reagents and colorimetric, fluorescent, or chemiluminescent readouts. Developed concurrently with advances in molecular biology exemplified by work at institutions such as Karolinska Institutet, Uppsala University, National Institutes of Health, and historical laboratories associated with figures like Niels Bohr and Marie Curie, the assay became a foundational tool in diagnostics across settings influenced by organizations such as the World Health Organization, Centers for Disease Control and Prevention, and Médecins Sans Frontières. Widely applied in contexts linked to events like the HIV/AIDS pandemic, the assay underpins testing strategies employed by agencies including Food and Drug Administration, European Medicines Agency, and Bill & Melinda Gates Foundation funded programs.
The assay emerged during a period of methodological innovation alongside contributions from researchers connected with Karolinska Institutet and Uppsala University and subsequent dissemination through conferences at venues such as Cold Spring Harbor Laboratory and Gordon Research Conferences. Early adopters included laboratories at the National Institutes of Health, hospitals associated with Mayo Clinic, Cleveland Clinic, and public health agencies like the Centers for Disease Control and Prevention, which integrated the method into surveillance programs for pathogens such as HIV-1, Hepatitis B virus, and Plasmodium falciparum. Commercialization and standardization were influenced by biotechnology firms inspired by breakthroughs reported in journals linked to publishers like Nature Publishing Group and Elsevier.
At core, the technique relies on antigen–antibody specificity and enzymatic signal amplification, principles shared with methodologies developed in parallel by researchers at institutions such as Harvard University, Stanford University, and University of Oxford. Common formats include direct, indirect, sandwich, and competitive ELISA, each adapted for targets ranging from proteins studied in laboratories at Salk Institute and Scripps Research to hormones measured in clinics affiliated with Johns Hopkins Hospital and Massachusetts General Hospital. Enzyme labels such as horseradish peroxidase and alkaline phosphatase became standard reagents marketed by companies like Thermo Fisher Scientific and Sigma-Aldrich, and detection systems evolved alongside instrumentation from manufacturers including Bio-Rad Laboratories and Agilent Technologies.
Typical consumables include microtiter plates produced by vendors associated with trade shows hosted by BIO International Convention and antibodies generated in facilities linked to European Molecular Biology Laboratory. Reagents often reference standards developed by organizations such as World Health Organization, National Institute of Standards and Technology, and assays are calibrated using reference materials circulated through networks like Global Polio Laboratory Network and programs supported by Gavi, the Vaccine Alliance. Enzyme substrates, blocking buffers, and wash solutions trace commercial supply chains connected to corporations such as Merck Group and BD (Becton, Dickinson and Company).
Typical protocols derive from methodological manuscripts disseminated via conferences at Cold Spring Harbor Laboratory and guidelines issued by agencies like Centers for Disease Control and Prevention and World Health Organization. Steps include plate coating with capture molecules, blocking to prevent nonspecific binding, incubation with samples and detection antibodies, and enzymatic development followed by plate reading on microplate readers from Tecan Group or PerkinElmer. Laboratories implementing high-throughput workflows often integrate automation platforms showcased at exhibitions such as Hannover Messe and collaborate with core facilities at institutions like Broad Institute and Wellcome Sanger Institute.
The assay is utilized for infectious disease diagnostics in programs led by World Health Organization and Centers for Disease Control and Prevention, seroepidemiology in studies performed at London School of Hygiene & Tropical Medicine and Imperial College London, vaccine immunogenicity measured in trials run by organizations such as Vaccine Alliance partners, and biomarker quantification in translational research at National Institutes of Health intramural centers. Clinical laboratories in hospitals including Massachusetts General Hospital, Johns Hopkins Hospital, and Ruth P. Ammon Medical Center apply the technique for hormone assays, allergen testing, and autoantibody panels, often within regulatory frameworks administered by Food and Drug Administration and European Medicines Agency.
Advantages—sensitivity, adaptability, and cost-effectiveness—contributed to adoption across academic centers like University of California, San Francisco and industry labs at Genentech. Limitations include cross-reactivity, matrix effects, and standardization challenges addressed in interlaboratory studies coordinated by entities such as International Organization for Standardization, Clinical and Laboratory Standards Institute, and consortiums comprising institutions like Harvard Medical School and Yale School of Medicine.
Quality assurance practices align with accreditation programs from College of American Pathologists, Joint Commission, and quality frameworks promoted by World Health Organization and Centers for Disease Control and Prevention. Interpretation of results often incorporates statistical methods taught in courses at London School of Economics and quality assessments reported in literature published by Oxford University Press and Cambridge University Press, with proficiency testing coordinated through networks like the European Centre for Disease Prevention and Control.
Category:Immunology