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By c rao

This quantity goals to give the elemental ideas, measurements, and methods of infrared spectroscopy and to survey, as thoroughly as attainable, its chemical purposes.

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Example text

Nsh = CVnsb 100 × Rnsb  (36) where: CVnsb is the coefficient of variation around the mean level of non-specific binding. Radioactivity, chemiluminescence and fluorescence signals follow a Poisson distribution. The total observed signal response in the absence of unlabeled antigen is the sum of the specific binding, Ro and that due to non-specific binding, Rnsb. Substitution of [B]/[F]det permits calculation of the total antigen concentration present (labeled and unlabeled) at the limit of sensitivity, and hence the sensitivity can be derived as: FIGURE 19 Antibody dilution curves.

If the sensitivity in the assay is 1 pmol/L but the sample is diluted 1/5 by the reagents, then it follows that the practical sensitivity is 5 pmol/L. These series of simple equations are very useful in enabling ‘what if’ calculations to be performed by varying the reactant concentrations and equilibrium constant. Theoretical dose–response curves may also be produced by substitution of the relevant parameters into Eqn (28) and solving the quadratic to obtain [B]/[F] and hence percentage bound. Figure 19 above shows the percentage binding of antigen by dilutions of antibody with an average equilibrium constant of 1 × 1010 L/mol.

E = CVe 100 × Ro (35) 42 The Immunoassay Handbook where: CVe is the coefficient of variation due to experimental errors. nsh = CVnsb 100 × Rnsb  (36) where: CVnsb is the coefficient of variation around the mean level of non-specific binding. Radioactivity, chemiluminescence and fluorescence signals follow a Poisson distribution. The total observed signal response in the absence of unlabeled antigen is the sum of the specific binding, Ro and that due to non-specific binding, Rnsb. Substitution of [B]/[F]det permits calculation of the total antigen concentration present (labeled and unlabeled) at the limit of sensitivity, and hence the sensitivity can be derived as: FIGURE 19 Antibody dilution curves.

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