Radioimmunoassay in Basic and Clinical Pharmacology

1 Radioimmunoassay: Historical Aspects and General Considerations.- A. Historical Aspects.- B. Principle, Practices, and Pitfalls.- I. Principle.- II. Practices.- III. Pitfalls.- C. Conclusions.- References.- 2 Basic Principles of Antigen-Antibody Interaction.- A. Introduction to the Immune System.- B. Antigens.- I. Chemical Nature.- II. Antigenic Determinants (Epitopes).- III. Haptens and Carriers.- IV. Size of Determinants.- V. Sequential and Conformational Epitopes.- C. Antibodies.- I. The Ab Combining Site (Paratope).- D. The Immune Interaction.- I. The Forces Involved.- II. Paratope-Epitope Fit.- III. Specificity.- IV. Affinity.- E. Effects of Ab–Ag Interaction.- I. Free Ag Binds Ig Receptor.- II. Free Ag Binds Free Ab.- III. Free Ab Binds Cell-Associated Ag.- F. Immune Interaction as a Signal: Role of Conformational Changes.- I. Antibody.- II. Antigen.- References.- 3 Production of Antisera by Conventional Techniques.- A. Introduction.- B. Substances Which Are Able to Evoke Immunogenic Responses Per Se.- C. Special Problems with Small Peptides.- D. Haptens Covalently Coupled to Protein Carriers.- I. Preparation of Derivatives Containing Reactive Groups.- II. Covalent Coupling to Protein Carriers.- E. Immune Response to Hapten-Protein Conjugates.- I. Effect of Method and Duration of Immunization.- II. Role of Adjuvants in Antibody Production.- III. Effect of Carrier Protein on Characteristics of Antisera.- IV. Effect of Hapten: Protein Molar Ratio on Characteristics of Antisera.- V. Effect of Site of Hapten Linkage to Protein on Characteristics of Antisera.- F. Characterization of Antisera.- I. Titer.- II. Affinity.- III. Sensitivity.- IV. Specificity.- References.- 4 Production of Monoclonal Antibodies for Radioimmunoassays.- A. Introduction.- I. Rationale for the Production of Monoclonal Antibodies.- II. Applications of Monoclonal Antibodies.- III. Problems.- B. Techniques.- I. Materials.- II. Methods.- C. Results and Outlook.- References.- 5 Radioiodination and Other Labeling Techniques.- A. Labeling for Immunologic Assay: Introduction.- B. Radioiodination.- I. General Considerations.- II. Chemistry.- C. Nonisotopic Labeling.- I. General Considerations.- II. Labeling with Enzymes.- III. Labeling with Fluorescent Compounds.- IV. Labeling with Luminescent Compounds.- V. Other Labeling Methods.- D. Conclusions.- References.- 6 Strategies for Developing Specific and Sensitive Hapten Radioimmunoassays.- A. Introduction.- B. Hapten-Carrier Conjugation.- I. Haptens with Amino Groups.- II. Haptens with Carboxyl Groups.- III. Haptens with Other Functional Groups.- C. Strategy for Increasing Specificity.- I. General Considerations: From Hapten Size to Epitope Size.- II. Steroids.- III. Other Small Haptens.- IV. Peptides.- V. Transformation of Immunogen.- D. Immunoassay Sensitization.- I. Increase in Tracer Specific Activity.- II. Modification of Reagent Concentrations and Assay Procedures..- E. Validation.- I. Nonantigenic Materials.- II. Antigen-like Materials.- III. Example of Validation: Peptide Radioimmunoassay.- References.- 7 How to Improve the Sensitivity of a Radioimmunoassay.- A. Introduction.- B. Basic Considerations.- C. The Influence of Labeled Tracer.- D. The Influence of Antiserum Dilution.- E. The Role of Incubation Volume.- F. Temperature and pH Effects.- G. Disequilibrium Conditions.- H. Other Factors Affecting Sensitivity.- References.- 8 Statistical Aspects of Radioimmunoassay.- A. Introduction.- B. The Logit-Log Method.- C. Is the Logit-Log Method Failing?.- D. The Four-Parameter Logistic.- E. Examples of Problems.- I. Evaluating Goodness of Fit.- II. Pooling of Information Over Assays.- F. Strategy to Deal with Failure of Logistic Models.- I. Does it Matter?.- II. Choice of Other Methods.- III. A “Universal” Approach.- G. Concluding Remarks.- References.- 9 Validation Criteria for Radioimmunoassay.- A. Introduction.- B. Conditions Necessary, but not Sufficient for Establishing the Validity of RIA Measurements.- C. Appropriate Biological Behaviour of the Measured Immunoreactivity.- D. Limited Cross-Reactions of Structurally Related Substances.- E. Use of Multiple Antisera.- F. Identical Chromatographic Behaviour of Standards and Unknowns.- G. Comparison with an Independent Method of Analysis.- H. Are we Measuring the Right Compound, in the Right Compartment?.- J. How to Evaluate an RIA Kit Critically?.- References.- 10 Measurement of Opioid Peptides in Biologic Fluids by Radioimmunoassay.- A. Introduction.- B. General Aspects.- I. Distribution and Processing of Opioid Peptides.- II. Characterization of Opioid Receptors.- C. Methodological Aspects.- I. Sampling Protocol.- II. Sample Workup (Preseparation).- III. Radioimmunoassay of Opioid Peptides from Individual Systems.- D. Functional Aspects.- I. Opioid Peptides in Biologic Fluids.- E. Clinical Aspects.- I. Pain.- II. Stress, Physical Exercise, and Shock.- III. Narcotic Dependence.- IV. Psychiatry.- V. Neuroendocrinology and Endocrine Tumors.- F. Concluding Remarks.- References.- 11 Radioimmunoassay of Pituitary and Hypothalamic Hormones.- A. Introduction.- B. General Principles of Determination.- I. The Antibody.- II. The Labeled Antigen.- III. Reference Preparations.- IV. Separation of Bound from Free Labeled Hormone.- V. Future Development of RIAs.- C. Radioimmunoassay of Anterior Pituitary Hormones.- I. Prolactin.- II. Growth Hormone.- III. Glycoprotein Hormones.- IV. Adrenocorticotropin, Endorphins, and Related Peptides.- D. Radioimmunoassay of Hypothalamic Hypophysiotropic Hormones.- I. Premises.- II. General Principles of Determination.- III. Features Peculiar to the Determination of a Single Hypophysiotropic Hormone.- IV. Significance of Hypophysiotropic Peptides in Biologic Fluids.- References.- 12 Radioimmunoassay of Nonpituitary Peptide Hormones.- A. Hormones Involved in Regulation of Carbohydrate Metabolism.- I. Insulin.- II. C-Peptide of Insulin.- III. Glucagon.- B. Hormones Regulating Calcium Homeostasis.- I. Physiology of Calcium Homeostasis.- II. Parathyroid Hormone.- III. Calcitonin.- References.- 13 Radioimmunoassay of Gastrointestinal Polypeptides.- A. Introduction.- B. Gastrin.- I. Physiologic Relevance.- II. Chemical Composition.- III. Characteristics of Antibodies and Labeled Peptides.- IV. Other Published Assays.- C. Cholecystokinin.- I. Published Radioimmunoassays.- D. Secretin.- I. Chemistry.- II. Antibody Production.- III. Radioiodination.- IV. Radioimmunoassay Procedure.- V. Characterization of Antibodies.- VI. Sample Preparation for Radioimmunoassay.- VII. Other Published Radioimmunoassays.- E. Somatostatin.- I. Antibody Production.- II. Radioiodination.- III. Sampling and Treatment of Serum.- IV. Serum Extraction.- V. Radioimmunoassay Procedure.- VI. Detection Limit.- VII. Other Published Radioimmunoassays.- F. Motilin.- I. Chemistry.- II. Antibody Production.- III. Radioiodination.- IV. Radioimmunoassay Procedure.- V. Characterization of Antibodies.- VI. Other Published Radioimmunoassays.- G. Gastric Inhibitory Peptide.- I. Antibody Production.- II. Characterization of Antisera.- III. Radioiodination.- IV. Radioimmunoassay Procedure.- V. Other Published Radioimmunoassays.- H. Neurotensin.- I. Chemistry.- II. Preparation of Antigen.- III. Immunization of Animals.- IV. Radioiodination.- V. Radioimmunoassay Procedure.- VI. Characterization of the Antibody.- VII. Stability of Neurotensin.- VIII. Other Published Radioimmunoassays.- J. Vasoactive Intestinal Polypeptide.- I. Chemistry.- II. Antibody Production.- III. Radioiodination.- IV. Radioimmunoassay Procedure.- V. Characterization of Antibodies.- VI. Sample Preparation for Radioimmunoassay.- VII. Other Published Radioimmunoassays.- K. Pancreatic Polypeptide and Peptide YY.- I. Pancreatic Polypeptide.- II. Peptide YY.- References.- 14 Radioimmunoassay of Atrial Peptide Blood and Tissue Levels.- A. Introduction.- B. Development of Atriopeptin Radioimmunoassay.- I. Immunization.- II. Iodination of Peptides.- III. Titering and Sensitivity.- C. Assay of Tissues and Plasma.- I. Atrial Extracts.- II. Plasma Immunoreactivity.- III. Brain Atriopeptin.- D. Processing of the Atriopeptin Prohormone.- E. Summary.- References.- 15 Immunochemical Methods for Adrenal and Gonadal Steroids.- A. Chemistry and Nomenclature.- B. Immunoassay Methods for Steroids.- I. Basic Principles.- II. Synthesis of the Immunogen Derivative.- III. The Radioactive Tracer.- IV. Sample Preparation Before Immunoassay.- V. Bound/Free Separation Systems.- VI. Monoclonal Antibodies to Steroids.- VII. Quality Assessment.- VIII. Alternative Immunoassay Methods.- C. The Adrenal Cortex.- I. Physiology.- II. Control Mechanisms for the Release of Cortisol.- III. Control Mechanisms for the Release of Aldosterone.- IV. Biosynthesis of Adrenal Steroids.- V. Metabolism of Adrenal Steroids.- VI. Immunochemical Methods for Adrenal Steroids.- D. The Testis.- I. Physiology.- II. Transport of Testosterone in Blood and Its Action at the Target Level.- III. Immunochemical Methods for Androgens.- E. The Ovary.- I. Physiology.- II. Immunochemical Methods for Ovarian Steroids.- References.- 16 Radioimmunoassay of Thyroid Hormones.- A. Introduction.- B. Iodometric Techniques.- I. PBI.- II. BEI.- III. T4Ic.- C. Radioligand Assays of Total T4 (TT4) and Total T3 (TT3).- I. TT4Assays.- II. TT3Assays.- III. Factors Affecting the Diagnostic Accuracy of Total Thyroid Hormone Determination.- D. Free Thyroid Hormone Assays.- I. Indirect Methods for the Estimation of Free Thyroid Hormone.- II. Direct Methods for Free Thyroid Hormone Determination.- III. Factors Affecting the Diagnostic Accuracy of Free Thyroid Hormone Measurements.- E. Radioimmunoassay of Other Iodothyronines, Iodotyrosines, and Products of Thyroid Hormone Degradation.- I. rT3RIA.- II. RIA of Diiodothyronines.- III. RIA of Monoiodothyronines.- IV. RIA of Tetraiodothyroacetic Acid (TETRAC) and Triiodothyroacetic Acid (TRIAC).- V. RIA of Iodotyrosines (MIT and DIT).- References.- 17 Radioimmunoassay of Catecholamines.- A. Introduction.- B. Radioimmunoassay of Catecholamines and Their Metabolites.- I. Antibodies to Catecholamines.- II. Antibodies to Metabolites of Catecholamines.- C. Summary.- References.- 18 Radioimmunoassay of Prostaglandins and Other Cyclooxygenase Products of Arachidonate Metabolism.- A. Introduction.- B. Biosynthesis and Metabolism of Prostaglandins and Thromboxanes.- C. Development of Radioimmunoassays for Prostaglandins and Thromboxanes.- I. Preparation of Immunogens.- II. Immunization.- III. Labeled Ligands.- IV. Separation of Antibody-Bound and Free Fractions of Ligands.- D. Validation of Radioimmunoassays for Cyclooxygenase Products of Arachidonic Acid Metabolism.- E. Factors that Affect the Validity of Prostaglandin and Thromboxane Radioimmunoassay Results.- I. Extraction and Purification Procedures.- II. The Blank Problem.- III. Problems Associated with the Determination of Tissue and Plasma Concentrations of Prostanoids.- F. Radioimmunoassay for Various Prostaglandins and Thromboxanes.- I. Prostaglandin F2?.- II. 15-Keto-13, 14-dihydro-prostaglandin F2?.- III. Prostaglandin E2.- IV. 15-Keto-l 3,14-dihydro-prostaglandin E2.- V. Prostaglandin E1.- VI. Prostaglandin D2.- VII. Indirect Determination of Prostaglandin Endoperoxides and Thromboxane A2.- VIII. Thromboxane B2 and 11-Dehydro-thromboxane B2.- IX. 15-Keto-13,14-dihydro-thromboxane B2.- X. 6-Keto-prostaglandin F1?.- XI. Metabolites of Prostaglandin I2 and 6-Keto-prostaglandin F1?.- XII. Main Urinary Metabolites of Prostaglandin F2? and Prostaglandin E2.- G. Comparison of Radioimmunoassay with Other Methods for Quantitative Determination of Cyclooxygenase Products of Arachidonic Acid Metabolism.- H. Radioimmunoassay of Cyclooxygenase Products of Arachidonic Acid Metabolism in Basic and Clinical Pharmacology.- J. Concluding Remarks.- References.- 19 Radioimmunoassay of Leukotrienes and Other Lipoxygenase Products of Arachidonate.- A. Introduction.- B. Biosynthesis and Metabolism of Lipoxygenase Products.- C. Biologic Activities of Lipoxygenase Products.- D. Biologic Assays to Measure Lipoxygenase Products.- E. Physicochemical Separation and Identification of Lipoxygenase Products.- F. Radioimmunoassays for Lipoxygenase Products.- I. Introduction.- II. Radioimmunoassays to Measure LTC4.- III. Radioimmunoassays to Measure LTD4.- IV. Radioimmunoassays to Measure LTB4.- V. Radioimmunoassays to Measure 12-HETE.- VI. Radioimmunoassays to Measure 15-HETE.- G. Sample Preparation for Radioimmunoassays.- H. Combination of Radioimmunoassays with Physicochemical Separation Methods.- J. Conclusions.- References.- 20 Radioimmunoassay of Cyclic Nucleotides.- A. Introduction.- B. Preparation of the Immunogen.- C. Immunization.- D. Preparation of the Radioindicator Molecule.- E. Evaluation of Antisera and Routine Immunoassay Procedures.- I. Double-Antibody Procedure.- II. Ammonium Sulfate Precipitation.- III. Charcoal.- F. Tissue Extraction.- G. Derivatization of Tissue Samples.- H. Pitfalls in Immunoassay Measurements.- J. Validation of the Immunoassay.- K. Special Applications of the Immunoassays.- References.- 21 Radioimmunoassay of Platelet Proteins.- A. Introduction.- B. Sample Collection and Processing.- C. Assay of ß-Thromboglobulin.- D. Assay of Platelet Factor 4.- E. Assay of Other Platelet Proteins.- F. Applications of Platelet-Specific Protein Radioimmunoassays.- G. Discussion.- References.- 22 A Competitive Binding Assay for Heparin, Heparan Sulphates and Other Sulphated Polymers.- A. Introduction.- B. Radiolabelling of Glycosaminoglycans.- I. Derivatisation.- II. Iodination.- C. Synthesis of Binding Reagent.- D. Assay for Therapeutic Heparins and Heparinoids.- I. Method.- II. Sensitivity.- III. Specificity.- IV. Studies in Human Volunteers.- E. Assay for Endogenous Heparan Sulphate.- I. Method.- II. Sensitivity.- III. Specificity.- IV. Results of Rat and Human Studies.- F. Other Applications.- G. Summary.- References.- 23 Radioimmunoassay of the Somatomedins/lnsulin-like Growth Factors.- A. Introduction and Nomenclature of the Somatomedins.- I. Methods for Measuring Somatomedins Before the Development of Radioimmunoassays.- B. Radioimmunoassay for Sm-C/IGF-I.- I. Antisera Production, Sensitivity, and Specificity.- II. Influence of Binding Proteins.- III. Clinical Utility of Measurements of Sm-C/IGF-I in Plasma.- IV. Heterologous Radioimmunoassays for Sm-C/IGF-I.- V. Measurement of Sm-C/IGF-I in Tissues and Media Extracts.- C. Radioimmunoassay for IGF-II.- References.- 24 Radioimmunoassay of Drugs and Neurotransmitters.- A. Introduction.- B. Methods for Coupling Neurotransmitters to Carrier Proteins.- I. Acetylcholine.- II. Serotonin.- III. Melatonin.- C. Methods for Coupling Drugs to Carrier Proteins.- I. Barbiturates.- II. Curare.- III. Clonidine.- IV. Phenothiazines.- V. Benzodiazepines.- VI. Butyrophenones.- VII. Atropine.- VIII. Opiates.- References.