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ELISPOT (Enzyme-Linked ImmunoSpot Assay, synonyms: Immunospot, ELISA Spot, ELISA-plaque) becomes an established tool in modern medicine. This technique has already a good standing in biotechnology (production and selection of antibodies), vaccinology (production and testing of vaccines), immunology (research and diagnosis), infectious diseases (sensitive detection of infections), and oncology (development of immunotherapeutic vaccines). There is also an increased use of the ELISPOT in experimental and clinical allergology. Similar to ELISA, ELISPOT is based on antibody pairs, including coating (catching) and detection antibodies. There are, however, substantial differences between these two techniques, e.g. the binding takes place in cell cultures. Thanks to this feature, ELISPOT overcomes a range of technical problems of ELISA and provides a high sensitivity. It is capable of detecting one single cell that secretes a protein of interest (a cytokine, effector protein, receptor, surface marker or an antibody), out of 1,000,000 cells. ELISPOT is not affected by the consumption of cytokines in the cell cultures, which is important advantage while detecting e.g. IL-4 or IL-2. A further development of the technique is Dual ELISPOT that allows the detection of more than one protein secreted by a cell. By combining ELISPOT with ELISA, a mean cytokine production by stimulated cells can be assessed. As ELISPOT is not yet widely known in Poland, the aim of this review is to present some details on the method, with its advantages and drawbacks. Also an overview of established and emerging applications of ELISPOT will be given in basic and clinical allergology and immunology. Key words: laboratory techniques, immunoenzymatic assays, ELISPOT, ELISA, immunology, allergology, oncology, vaccinology. |
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© Radoslaw Spiewak
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