The ELISA, enzyme-linked immunosorbent assay

The Burnham Institute, La Jolla, CA, USA.
Clinical Chemistry (Impact Factor: 7.91). 10/2009; 56(2):319-20. DOI: 10.1373/clinchem.2009.127803
Source: PubMed
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    • "ELISA is also based on antibody-binding, but unlike RIA, the read-out is based on an enzymatic reaction (e.g. horseradish peroxidase) coupled to one of the reagents and resulting in a colorimetric or otherwise traceable signal (Engvall & Perlmann, 1971). The detection of the enzymatic conversion of a suitable substrate to a detectable metabolite is often based on simple spectrophotometry (typically at 450 nm), but other detection systems (chemiluminescence, electrochemiluminescence and fluorescence) exist. "
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    ABSTRACT: The two incretin hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP), are secreted from the gastrointestinal tract in response to meals and contribute to the regulation of glucose homeostasis by increasing insulin secretion. Assessment of plasma concentrations of GLP-1 and GIP is often an important endpoint in both clinical and preclinical studies and, therefore, accurate measurement of these hormones is important. Here, we provide an overview of current approaches for the measurement of the incretin hormones, with particular focus on immunological methods.
    Full-text · Article · Dec 2014 · Journal of Diabetes and its Complications
    • "non-specific binding of plasma proteins to blocked plates with no factor H coat) is suggested instead of using only a reference well or blank well subtraction approach. Since Engvall and Perlmann established ELISA in 1971 (Engvall and Perlmann, 1971), it has become one of the most powerful diagnostic tools in medical research and diagnosis. The ELISA has many steps that require optimisation, with coat concentration, block, wash and secondary specificity being generally regarded as central to obtaining the maximal signal to noise ratio. "
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    ABSTRACT: The screening of all atypical haemolytic uraemic syndrome (aHUS) patients for factor H autoantibodies is best practice. However, there is no consensus assay for the reporting of factor H autoantibody titres. In this study, three European complement laboratories with expertise in the field of autoantibody testing address this by systematically evaluating several ELISA methods used for the detection of factor H autoantibodies. All methods tested adequately detect high titre samples. However, this study recommends the Paris method for the detection and reporting of factor H autoantibodies to be used when setting up a factor H autoantibody screen. The importance of individual sample background subtraction in these ELISA tests was established. The use of a relative or arbitrary unit index with a common positive and negative serum allowed for consistent comparison of findings from different test centres. Therefore, it is recommended that a standard arbitrary unit scale based on a titration curve from a common positive anti-serum be adopted to allow future establishment of the relative importance of particular titres of factor H autoantibodies in aHUS. Systematic assay for the presence of factor H autoantibodies in patients using the Paris method will provide the longitudinal analysis needed to fully establish the importance of factor H autoantibodies in disease. This will feed into additional research to clarify whether additional factors have a bearing on the phenotype/outcome of autoimmune aHUS.
    No preview · Article · Jun 2013 · Immunobiology
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    • "Transcription factors typically exist in the cell in an inactive state, and become activated by the presence of a specific ligand, leading to the expression of target gene(s). The inhibition or undesired activation of transcription factors can lead to a number of diseases including cancer, developmental disorders, abnormal hormone responses, autoimmunity and inflammation (5–10). The cellular levels of such proteins can thus be used as diagnostic markers for many diseases, and the targeting of transcription factor activity represents a potential avenue for therapeutic intervention (11–13). "
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    ABSTRACT: Transcription factors play a central role in cell development, differentiation and growth in biological systems due to their ability to regulate gene expression by binding to specific DNA sequences within the nucleus. The dysregulation of transcription factor signaling has been implicated in the pathogenesis of a number of cancers, developmental disorders, inflammation and autoimmunity. There is thus a high demand for convenient high-throughput methodologies able to detect sequence-specific DNA-binding proteins and monitor their DNA-binding activities. Traditional approaches for protein detection include gel mobility shift assays, DNA footprinting and enzyme-linked immunosorbent assays (ELISAs) which tend to be tedious, time-consuming, and may necessitate the use of radiographic labeling. By contrast, luminescence technologies offer the potential for rapid, sensitive and low-cost detection that are amenable to high-throughput and real-time analysis. The discoveries of molecular beacons and aptamers have spear-headed the development of new luminescent methodologies for the detection of proteins over the last decade. We survey here recent advances in the development of luminescent detection methods for DNA-binding proteins, including those based on molecular beacons, aptamer beacons, label-free techniques and exonuclease protection.
    Full-text · Article · Oct 2011 · Nucleic Acids Research
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