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Buchumschlag
Gespeichert in:
Bibliographische Detailangaben
Weitere beteiligte Personen: Gosling, James Patrick (HerausgeberIn)
Format: Buch
Sprache:Englisch
Veröffentlicht: New York, NY Oxford University Press 2005
Ausgabe:Repr.
Schriftenreihe:The practical approach series
Schlagwörter:
Methode
Immunassay
Links:http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=015660321&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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Umfang:XIX, 304 S. Ill., graph. Darst.
ISBN:0199637105
Internformat

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Datensatz im Suchindex

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adam_text Immunoassays is the first practical manual designed to help any biologist develop an immunoassay in any common format for any suitable analyte. The book starts by describing the basic principles of immunoassay and by evaluating the choice of assay types and formats. Following chapters describe, in full practical detail: methods for raising and choosing polyclonal and monoclonal antibodies, immunogen preparation and characterization, sourcing standards, the synthesis and characterization of radioactive, enzymatic, fluorescent and ligand labels, the preparation of a wide variety of solid-phase reagents, assay optimization (sensitivity and accuracy), validation of quantitative and qualitative assays, curve fitting and data processing (both manual and computer based), and quality assurance proce- dures for small and large scale users. This volume is unique in its scope and coverage of immunoassays. It is highly recommended both for researchers new to immunoassays and those who are seek- ing an updated source guide. The mix of background information, step by step protocols, and strong practical advice for achieving success will prove invaluable to any life scientist who chooses to use immunoassays in either a research environment or for routine testing. Cover illustration: Marcus Gosling OXFORD UNIVERSITY PRESS ISBN 0-19-963710-5 78 199 6371 www.oup.com  Contents Preface v Protocol List xiii Abbreviations xvii 1 Analysis by specific binding i James P. Gosling 1 Introduction 1 2 Requirements for high-affinity specific binding 2 immunoglobulins 2 Antigen-antibody binding 5 3 The variety of immunoassays 7 Representing assay complexes in text 7 Classification 8 Nomenclature 23 The variety of formats 24 4 The limitations of binding assays 14 2 Raising antibodies 29 Marian M. Kane and John N. Banks 1 Introduction 19 Monoclonal and polyclonal antibodies 29 Antibodies and immunoassays 20 2 Animals and cell lines 21 Raising monoclonal antibodies 21 Raising polyclonal antibodies 23 3 Immunization 25 The immunogen 25 Adjuvants 26 Injection route and timing 29 4 Monitoring immunizations 32 General and when raising polyclonal antisera 32 When raising monoclonal antibodies 33 Screening procedures 33 CONTENTS 5 Making and manipulating hybridoma cells 37 Facilities and media 37 Fusions and selection of hybridomas 41 Cloning and further selections 45 Maintenance and storage 47 The production of monoclonal antibodies 49 6 Antibody purification 51 7 Emerging technologies 55 In vitro immunization 55 Recombinant antibody techniques 55 Choosing and characterizing antibodies 59 Anthony Forde and John Coley 1 Introduction 59 Secondary antibodies 59 Specificity 61 Epitope mapping 61 Affinity 62 2 Sourcing antibodies 67 3 Exploring specificity 69 Specificity as assessed by suppliers 69 Specificity and the assay developer 70 4 Estimating affinity 76 Approaches 76 Methods 77 5 Conclusion 86 4 Labels and endpoints 89 Wajdi Abdul-Ahad and Martin Brett 1 Labelling substances 89 Radioisotopic labels 89 Enzyme labels 92 Fluorescent labels 93 Ligands and indirect labelling 94 2 Enzyme measurement methods 95 Methods with chromogens 95 Methods with fluorogens 97 Methods that generate luminescence 98 3 Labelling and conjugation 99 Radiolabelling 99 The chemistry of conjugation 104 Protein-protein conjugation 109 Enzyme hapten coupling 117 Fluorescence labelling 121 Biotin coupling 122 4 Label characterization 123 5 Conjugate stabilization 125 viii S Solid-phase reagents 129 Robert S. Matson 1 Introduction 129 Liquid-phase assay with solid-phase separation 130 Choice of solid-phase support 130 2 Approaches to immobilization 131 Preparation of ligands for immobilization 132 Covalent attachment 132 Noncovalent adsorption 140 Indirect immobilization 141 Treatments after coating 142 3 Solid-phase formats 144 Microtitre plates 146 Tubes 150 Membranes 150 Microspheres 155 4 Handling and storage 160 Surface dénaturation 160 Microtitre wells and tubes 160 Membranes 161 Micro spheres 161 5 Sourcing of reagents 161 6 Standards and Immunogens 165 M. Quasim Chaudry 1 Introduction 165 Criteria for a good immunogen 165 Animal welfare 167 2 Purification of immunogens 168 3 Synthetic and recombinant immunogens 169 Synthetic peptide immunogens 169 Recombinant peptide immunogens 170 4 Conjugation of haptens and carriers 170 Materials and reagents 171 Methods 172 Characterization of hapten immunogens 180 5. Storage of immunogens 182 6. Standards 182 7 Optimization 187 Eric Ezan and Jacques Grassi 1 Introduction 187 2 Sensitivity 187 Estimators of sensitivity 187 Optimization of reagent concentrations 191 Optimization of physiochemical parameters 194 CONTENTS 3 Accuracy 196 Factors affecting accuracy 196 Technological variability 298 Crossreactive compounds 199 Plasma-binding proteins 200 Anti-Ig antibodies 203 4 Conclusions 208 Validation 211 Peter O Fegan 1 Introduction 211 Validation in the development cycle 211 Validation criteria and analytical goals 212 2 Specificity 212 Crossreactivity studies 213 3 Precision 217 Materials 217 Levels of precision 218 4 Sensitivity 222 5 Accuracy 224 Recovery of added standard 224 Independence of volume 226 Comparison with a reference method 228 6 Biological validation 230 Concentrations and changes in concentration 230 Normal ranges and diagnostic cut-off levels 231 7 Qualitative and semi-quantitative tests 232 Specificity 232 Validation criteria 233 8 Evaluation of reagent stability 235 9 Data processing 239 Barry Nix and David Wild 1 Introduction 239 2 Fitting calibration curves 240 Definitions 240 Manual and computer methods 241 Computer methods in common use 245 Evaluating candidate methods 247 3 More worked examples 254 A good logit-log fit 254 Consequences of a poor logit-log fit 254 4 Calibration curve-fit software 260 X CONTENTS 10 Quality assurance 263 John Seth 1 Introduction 263 2 Quality assurance 263 Principles 263 Specimen collection and preservation 264 Reporting results 265 3 Internal quality control 265 Definitions 265 Pre-analytical quality control 266 Quality control of the analysis 268 Post analytical quality control 282 Limitation of IQC 282 4 External quality assessment 282 Principles 282 Uses and availability of EQA schemes 283 Operation of EQA schemes for immunoassay 285 Limitations of EQA 289 Making use of EQA and IQC data 292 A1 Suitable reagents and other requisites Index 299 XI
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series2 The practical approach series
spellingShingle Immunoassays a practical approach
Methode (DE-588)4038971-6 gnd
Immunassay (DE-588)4113998-7 gnd
subject_GND (DE-588)4038971-6
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title Immunoassays a practical approach
title_auth Immunoassays a practical approach
title_exact_search Immunoassays a practical approach
title_full Immunoassays a practical approach ed. by James P. Gosling
title_fullStr Immunoassays a practical approach ed. by James P. Gosling
title_full_unstemmed Immunoassays a practical approach ed. by James P. Gosling
title_short Immunoassays
title_sort immunoassays a practical approach
title_sub a practical approach
topic Methode (DE-588)4038971-6 gnd
Immunassay (DE-588)4113998-7 gnd
topic_facet Methode
Immunassay
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