Radioimmunoassay in Basic and Clinical Pharmacology by L. BartalenaRadioimmunoassay in Basic and Clinical Pharmacology by L. Bartalena

Radioimmunoassay in Basic and Clinical Pharmacology

byL. BartalenaEditorCarlo Patrono, Bernhard A. Peskar

Paperback | November 18, 2011

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Thirty years have elapsed since the first description by S. A. BERSON and R. S. Y ALOW of the basic principles of radioimmunoassay (RIA). During this period of time, RIA methodology has been instrumental to the growth of many areas of biomedical research, including endocrinology, oncology, hematology, and pharmacology. It has done so by providing a relatively simple universal tool allowing, for the first time, the detection of endogenous mediators that are present 12 10 in body fluids at concentrations as low as 10- _10- M. The fundamental nature of this discovery and the wide-ranging fall-out of basic and clinical knowledge derived from its application have been acknowledged by the many honors tributed to its pioneers, including the Nobel Prize awarded to Dr. Y ALOW 10 years ago. Although several excellent books have been published during the past decades covering various aspects of RIA methodology, we felt the need, as pharmacologists, for a comprehensive discussion of the methodological and conceptual issues related to the main classes of mediators of drug action and to drugs themselves. Thus, we gladly accepted the challenge provided by the invitation to edit a volume of the Handbook of Experimental Pharmacology on Radioimmunoassay in Basic and Clinical Pharmacology. We tried to balance the emphasis placed on more general aspects of the RIA methodology and that on specific mediators.
Title:Radioimmunoassay in Basic and Clinical PharmacologyFormat:PaperbackDimensions:611 pages, 24.4 × 17 × 1.73 inPublished:November 18, 2011Publisher:Springer-Verlag/Sci-Tech/TradeLanguage:English

The following ISBNs are associated with this title:

ISBN - 10:3642718116

ISBN - 13:9783642718113

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Table of Contents

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.