title: ELISPOT Assay for Measurement of Antigen-Specific and Polyclonal Antibody Responses.
abstract: The enzyme-linked immunospot (ELISPOT) assay for detection of antigen-specific and polyclonal antibody responses by single antibody-secreting cells has become the method of choice due to its cell-based quantitative value. Antigen stability and specificity and the diversity of antigens that can be used in the assay have contributed to the translational application of ELISPOT as demonstrated by many FDA-approved clinical tests that employ this technique. In addition, the ELISPOT assay can be used to detect two antigenically different secreted antibodies simultaneously by two-color analysis and offers the unique possibility of quantifying the number of antibody molecules secreted per cell.
authors: Department of Microbiology and Immunology, University of Göteborg, Göteborg, Sweden.
authors: Nils
authors: N
authors: Lycke
authors: Department of Cell Biology, SUNY Downstate College of Medicine, Brooklyn, New York.
authors: Richard
authors: R
authors: Coico
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000921:Antibody-Producing Cells / Q000276:immunology / Q000378:metabolism; D000941:Antigens / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay / Q000295:instrumentation / Q000379:methods; D000939:Epitopes / Q000276:immunology*; D006801:Humans
chemical_list: D000906:Antibodies; D000941:Antigens; D000939:Epitopes
keywords: ELISA; ELISPOT; antibody-secreting cells
references: Alexander, N. , Fox, A. , Lien, V.T. , Dong, T. , Lee, L.Y. , Hang , Nle, K. , Mai, le Q. , and Horby, P. 2013. Defining ELISpot cut-offs from unreplicated test and control wells. J. Immunol. Methods 392:57-62.
references: Bland, M.J. and Altman, D.G. 1986. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1:307-311.
references: Czerkinsky, C. , Nilsson, L.Å. , Nygren, H. , Ouchterlony, Ö. , and Tarkowski, A. 1983. A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J. Immunol. Methods 65:109-121.
references: Czerkinsky, C. , Moldeveanu, Z. , Mestecky, J. , Nilsson, L.Å. , and Ouchterlony, Ö. 1988a. A novel two-color ELISPOT assay. I. Simultaneous detection of distinct types of antibody-secreting cells. J. Immunol. Methods 115:31-37.
references: Czerkinsky, C. , Nilsson, L.Å. , Tarkowski, A. , Koopman, W.J. , Mestecky, J. , and Ouchterlony, Ö. 1988b. The enzyme-linked immunospot assay for detection of specific antibody-secreting cells: Methodology and applicability. In Theoretical and Technical Aspects of ELISA and Other Solid Phase Immunoassays ( D.M. Kemeny and S.J. Challacombe , eds.) pp. 217-239. John Wiley & Sons, New York.
references: Jahnmatz, M. , Kesa, G. , Netterlid, E. , Buisman, A. , Thorstensson, R. , and Ahlborg, N. 2013. Optimization of a human IgG B-cell ELISpot assay for the analysis of vaccine-induced B-cell responses. J. Immunol. Methods 391:50-59.
references: Franci, C. , Ingles, J. , Castro, R. , and Vidal, J. 1986. Further studies on the ELISA-spot technique. Its application to particulate antigens and potential improvement in sensitivity. J. Immunol. Methods 88:225-232.
references: Lycke, N. 1986. A sensitive method for the detection of specific antibody production in different isotypes from single lamina propria plasma cells. Scand. J. Immunol. 24:393-403.
references: Lycke, N. , Kilander, A. , Nilsson, L.Å. , Tarkowski, A. , and Werner, N. 1989. Production of antibodies to gliadin in intestinal mucosa of patients with coeliac disease: A study at the single cell level. Gut 30:72-77.
references: Möller, S.A. and Borrebaeck, C.A.K. 1985. A filter immuno-plaque assay for the detection of antibody-secreting cells in vitro. J. Immunol. Methods 79:195-204.
references: Sedgwick, J.D. and Holt, P.G. 1983. A solid-phase immunoenzymatic technique for the enumeration of specific antibody-secreting cells. J. Immunol. Methods 57:301-309.