introduction to radioimmunoassay and related techniques

(Subsections not included) Chapter 1: 1.1 Introduction. 1.2 Terminology. 1.3 Early development of radioimmunoassay. 1.4 Basic principles of binding assays. 1.5 Binder dilution curves and standard curves. 1.6 Methods for plotting the standard curve. 1.7 The measurement of K value. Chapter 2: 2.1 The need for purified ligand. 2.2 Availability of pure ligand. 2.3 Dissimilarity between purified ligand and endogenous ligand. 2.4 Standards. 2.5 Storage of materials used in binding assays. Chapter 3: 3.1 Radioactive isotopes. 3.2 Counting of radioactive isotopes. 3.3 Choice of counter. 3.4 Some practical aspects of isotope counting. 3.5 Essential characteristics of a tracer. 3.6 Preparation of tracers. 3.7 Iodinated tracers. 3.8 Variations on the use of radiolabelled tracers. Chapter 4: 4.1 Particle labels. 4.2 Enzyme labels (enzymoimmunoassay, EIA). 4.3 Fluorescent labels (fluoroimmunoassay, FIA). 4.4 Luminescent labels. 4.5 Advantages and disadvantages of non-isotopic labels in immunoassays. 4.6 Conclusions: the place of non-isotopic binding assays. Chapter 5: 5.1 Antibodies and the immune response. 5.2 Preparation of Antisera for use in RIA. 5.3 'Monoclonal' antibodies. 5.4 Cell receptors. 5.5 Circulating binding proteins. 5.6 Assays for the detection of endogenous antibodies, circulating binding proteins and receptors. Chapter 6: 6.1 Efficiency of separation procedures. 6.2 Practicality of separation procedures. 6.3 Methods of separation of bound and free ligand. 6.4 Immunometric techniques. Chapter 7: 7.1 General aspects of extraction procedures. 7.2 Extraction using particulate adsorbents. 7.3 Extraction with immunoadsorbents. 7.4 Extraction with organic solvents. 7.5 Dissociation procedures. 7.6 Measurement of 'free' hormone or drug. 7.7 Conclusions - the elimination of extraction procedures. 7.8 Sample collection and transport for immunoassay. Chapter 8: 8.1 Calculation of results by manual extrapolation. 8.2 Data transformation of the standard curve. 8.3 The logit transformation. 8.4 Identification of outliers. 8.5 Estimation of confidence limits to the result of an unknown. 8.6 Computer calculation of results. 8.7 Calculation of results of labelled antibody assays. 8.8 Presentation of results. Chapter 9: 9.1 Definition of sensitivity. 9.2 Methods of increasing the sensitivity of a labelled antigen immunoassay. 9.3 Methods of increasing the sensitivity of a labelled antibody assay (immunometric assays). 9.4 Methods of decreasing the sensitivity of an assay. 9.5 The low-dose hook effect. 9.6 Targeting of binding assays - the importance of ranges. 9.7 Optimisation of an assay by theoretical analysis. 9.8 Conclusions. Chapter 10: 10.1 Definition of specificity. 10.2 Specific non-specificity. 10.3 Non-specific non-specificity. Chapter 11: 11.1 Definitions. 11.2 Factors affecting precision. 11.3 Quality control to monitor the precision of a binding assay. 11.4 Practical use of a quality-control scheme. 11.5 External quality control schemes.