Radioimmunoassay, enzyme and non-enzyme-based immunoassays

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Radioimmunoassay, enzyme and non-enzyme-based immunoassays R.D. Grange, J.P. Thompson, D.G. Lambert British Journal of Anaesthesia Volume 112, Issue 2, Pages 213-216 (February 2014) DOI: 10.1093/bja/aet293 Copyright © 2014 The Author(s) Terms and Conditions

Fig 1 (a) Sample peptide is incubated with primary antibody. (b) Radiolabelled peptide is then added. It competes with sample peptide and displaces it. (c) Secondary antibody binds to primary antibody and causes it to precipitate out of solution. (d) Centrifugation causes the antibody–antigen complex to form a pellet. (f) Example of a typical standard curve. (g) Actual standard curve for urotensin-II (UII) where amount of radioactive iodine bound is expressed as B/B0 which is the ratio of binding at each standard concentration, B to that bound in the absence of displacer, B0. Analyte samples in biological specimens should lie on the straight part of the curve. British Journal of Anaesthesia 2014 112, 213-216DOI: (10.1093/bja/aet293) Copyright © 2014 The Author(s) Terms and Conditions

Fig 2 Schematic showing the differences between direct (a), indirect (b), sandwich (c), and competitive (d) EIA methods. (e) Actual standard curve for a sandwich TNF-α assay. Note the way the standard curve is presented varies with the RIA in Figure 1, but analyte samples in biological specimens should lie on the straight part of the curve. RLU, relative light units signal from the enzyme reaction. British Journal of Anaesthesia 2014 112, 213-216DOI: (10.1093/bja/aet293) Copyright © 2014 The Author(s) Terms and Conditions