Rupprecht / Nagarajan Current Laboratory Techniques in Rabies Diagnosis, Research and Prevention, Volume 1

1. Auflage 2014
ISBN: 978-0-12-800465-4
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, 350 Seiten

Laboratory Techniques in Rabies Diagnosis, Research and Prevention provides a basic understanding of the current trends in rabies. It establishes a new facility for rabies surveillance, vaccine and antibody manufacturing. It offers clarity about the choice of laboratory methods for diagnosis and virus typing, of systems for producing monoclonal and polyclonal antibodies and of methods for testing potency of vaccines and antibodies. The book covers advancements in the classical methods described as well as recent methods and approaches pertaining to rabies diagnosis and research. - Supplies techniques pertaining to rabies diagnosis and research - Provides an update on the conventional and modern vaccines for rabies prevention - Offers updates on the full length antibodies and antibody fragments for post exposure prophylaxis of rabies - Presents technique descriptions that can be used to be compared to industry protocols to identify and establish potential new techniques

Autoren/Hrsg.

Rupprecht, Charles

Nagarajan, Thirumeni

Inhaltsverzeichnis

1;Front Cover;1
2;Current Laboratory Techniques in Rabies Diagnosis, Research and Prevention;4
3;Copyright Page;5
4;Contents;6
5;Foreword;14
6;List of Contributors;16
7;ONE: INTRODUCTION;20
7.1;1. Basic Facts about Lyssaviruses;22
7.1.1;1.1 Introduction;22
7.1.2;1.2 Virion and Genome Organization;23
7.1.3;1.3 Phylogeny and Serologic Cross-Reactivity of Lyssaviruses;25
7.1.4;1.4 Host Range;31
7.1.5;1.5 Pathobiology;34
7.1.6;References;36
8;TWO: RABIES DIAGNOSIS;42
8.1;A: Demonstration of Viral Subunits and Antigens;44
8.1.1;2. Demonstration of Lyssavirus Antigens by a Direct Rapid Immunohistochemical Test;46
8.1.1.1;2.1 Introduction;46
8.1.1.2;2.2 Materials;48
8.1.1.2.1;2.2.1 Reagents;48
8.1.1.2.2;2.2.2 Equipment;48
8.1.1.2.3;2.2.3 Biological Material: Deterioration or Decomposition;48
8.1.1.3;2.3 Methods;49
8.1.1.3.1;2.3.1 Performing Diagnostic Assay;49
8.1.1.3.2;2.3.2 Reading and Recording of Results;51
8.1.1.3.3;2.3.3 Test Results are Reported if the Following Observations are Made;52
8.1.1.4;2.4 Discussion;52
8.1.1.5;References;55
8.1.2;3. Demonstration of Rabies Virus Antigens by a Latex Agglutination Test;56
8.1.2.1;3.1 Introduction;56
8.1.2.2;3.2 Materials;57
8.1.2.2.1;3.2.1 Reagents;57
8.1.2.2.2;3.2.2 Biological Materials;57
8.1.2.2.3;3.2.3 Equipment;57
8.1.2.3;3.3 Methods;57
8.1.2.3.1;3.3.1 Production of Anti-Rabies Serum;57
8.1.2.3.2;3.3.2 Preparation of Anti-Rabies IgG;58
8.1.2.3.3;3.3.3 Latex Coating;58
8.1.2.3.4;3.3.4 Test Saliva;58
8.1.2.3.5;3.3.5 Slide Agglutination;58
8.1.2.4;3.4 Discussion;59
8.1.2.4.1;3.4.1 Experimental Tips;59
8.1.2.4.2;3.4.2 Technical Notes;59
8.1.2.4.3;3.4.3 Critical Parameters;60
8.1.2.5;References;61
8.1.3;4. Rabies Diagnosis: Demonstration of Viral Antigens by Flow Cytometry;62
8.1.3.1;4.1 Introduction;62
8.1.3.2;4.2 Materials;63
8.1.3.2.1;4.2.1 Reagents;63
8.1.3.2.2;4.2.2 Equipment;64
8.1.3.2.3;4.2.3 Biological Materials;64
8.1.3.3;4.3 Methods;64
8.1.3.3.1;4.3.1 Infection and Incubation;64
8.1.3.3.2;4.3.2 Cell Staining;64
8.1.3.3.3;4.3.3 Flow Cytometric Analysis of Infection;65
8.1.3.3.4;4.3.4 Interpretation of Results;65
8.1.3.4;4.4 Discussion;66
8.1.3.4.1;4.4.1 Experimental Tips;67
8.1.3.4.2;4.4.2 Critical Parameters and Troubleshooting;67
8.1.3.4.3;4.4.3 Precautions;68
8.1.3.4.4;4.4.4 Alternative Materials and Methods;68
8.1.3.4.5;4.4.5 Time Considerations;68
8.1.3.4.6;4.4.6 Limitations;69
8.1.3.4.7;4.4.7 Future Considerations;69
8.1.3.5;Acknowledgments;69
8.1.3.6;References;70
8.1.4;5. Demonstration of Rabies Virus Antigens by an Immunochromatographic Strip Test;72
8.1.4.1;5.1 Introduction;72
8.1.4.2;5.2 Materials;73
8.1.4.2.1;5.2.1 Reagents;73
8.1.4.2.2;5.2.2 Biological Materials;73
8.1.4.2.3;5.2.3 Materials and Equipment;74
8.1.4.3;5.3 Methods;74
8.1.4.3.1;5.3.1 Colloidal Gold Conjugation;74
8.1.4.3.2;5.3.2 Test Strip Preparation;74
8.1.4.3.3;5.3.3 Application of the Test Strip;75
8.1.4.4;5.4 Discussion;76
8.1.4.4.1;5.4.1 Experimental Tips;76
8.1.4.4.2;5.4.2 Technical Notes;76
8.1.4.4.3;5.4.3 Future Considerations;77
8.1.4.5;References;78
8.2;B: Demonstration of Viral Nucleic Acids;80
8.2.1;6. Demonstration of African Lyssavirus RNA with Real-Time Polymerase Chain Reaction;82
8.2.1.1;6.1 Introduction;82
8.2.1.2;6.2 Materials;83
8.2.1.2.1;6.2.1 Reagents;83
8.2.1.2.2;6.2.2 Equipment;83
8.2.1.2.3;6.2.3 Biological Material;84
8.2.1.3;6.3 Methods;84
8.2.1.3.1;6.3.1 Preparation of Reagent Master Mix;84
8.2.1.3.2;6.3.2 Cycling Protocol;85
8.2.1.3.2.1;6.3.2.1 Analysis of Results;86
8.2.1.4;6.4 Discussion;87
8.2.1.5;References;91
8.2.2;7. Real-Time Quantitative Polymerase Chain Reaction for the Demonstration of Lyssavirus Nucleic Acid;94
8.2.2.1;7.1 Real-Time Quantitative Polymerase Chain Reaction for the Demonstration of Lyssavirus Nucleic Acid;95
8.2.2.2;7.2 Methodology;95
8.2.2.2.1;7.2.1 Viral RNA Extraction;95
8.2.2.2.2;7.2.2 Generic Pan Lyssavirus Real-Time Polymerase Chain Reactions;95
8.2.2.2.3;7.2.3 Real-Time Polymerase Chain Reaction for Lyssavirus Species using Specific Taqman Probes;99
8.2.2.3;7.3 Discussion;101
8.2.2.4;Acknowledgments;102
8.2.2.5;References;102
8.2.3;8. Reverse Transcription-Loop-Mediated Isothermal Amplification System for the Detection of Rabies Virus;104
8.2.3.1;8.1 Introduction;104
8.2.3.2;8.2 Materials;105
8.2.3.2.1;8.2.1 Reagents;105
8.2.3.2.2;8.2.2 Primer Set for Reverse Transcription-Loop-Mediated Isothermal Amplification;105
8.2.3.2.3;8.2.3 Equipment;108
8.2.3.2.4;8.2.4 Biological Materials and Sample Preparation;109
8.2.3.3;8.3 Methods;109
8.2.3.3.1;8.3.1 Preparation of Mastermix;109
8.2.3.3.2;8.3.2 Mixing of Mastermix and Sample Solution;110
8.2.3.3.3;8.3.3 Amplification Reaction;110
8.2.3.3.4;8.3.4 Detection and Interpretation of Results;110
8.2.3.4;8.4 Discussion;112
8.2.3.5;Acknowledgments;113
8.2.3.6;References;113
8.2.4;9. Detection of Viral Nucleic Acids by In Situ Hybridization;116
8.2.4.1;9.1 Introduction;116
8.2.4.2;9.2 Materials;117
8.2.4.2.1;9.2.1 Reagents;117
8.2.4.2.2;9.2.2 Equipment;119
8.2.4.2.3;9.2.3 Biological Materials;119
8.2.4.2.4;9.2.4 Laboratory Animals;119
8.2.4.3;9.3 Methods;119
8.2.4.3.1;9.3.1 Generation of Control Tissues;119
8.2.4.3.2;9.3.2 Section Preparation;119
8.2.4.3.3;9.3.3 Production and Evaluation of Probe;120
8.2.4.3.4;9.3.4 In Situ Hybridization;121
8.2.4.3.5;9.3.5 Interpretation of Results;122
8.2.4.4;9.4 Discussion;122
8.2.4.4.1;9.4.1 Experimental Tips;122
8.2.4.4.2;9.4.2 Critical Parameters, Troubleshooting, and Precautions;123
8.2.4.4.3;9.4.3 Alternative Materials and/or Methods;123
8.2.4.4.4;9.4.4 Time Considerations;123
8.2.4.4.5;9.4.5 Limitations;124
8.2.4.4.6;9.4.6 Future Considerations;124
8.2.4.5;Acknowledgments;124
8.2.4.6;References;125
8.2.5;10. Genetic Characterization via Pyrosequencing;126
8.2.5.1;10.1 Introduction;126
8.2.5.2;10.2 Materials;127
8.2.5.2.1;10.2.1 Reagents and Equipment;127
8.2.5.2.2;10.2.2 Diagnostic Samples;128
8.2.5.3;10.3 Methods;128
8.2.5.3.1;10.3.1 Preliminary Phase – Population of the Local Database (IdentiFire Library);128
8.2.5.3.2;10.3.2 Phase 1 – RNA Extraction, One-Step RT-PCR, Gel Electrophoresis Analysis;128
8.2.5.3.3;10.3.3 Phase 2 – Pyrosequencing Reaction and Result Analysis;129
8.2.5.4;10.4 Discussion;130
8.2.5.4.1;10.4.1 Experimental Tips;130
8.2.5.4.2;10.4.2 Critical Parameters and Troubleshooting;130
8.2.5.4.3;10.4.3 Precautions;132
8.2.5.4.4;10.4.4 Alternative Methods;132
8.2.5.4.5;10.4.5 Time Consideration;133
8.2.5.4.6;10.4.6 Limitations;133
8.2.5.4.7;10.4.7 Future Considerations;134
8.2.5.5;Acknowledgments;134
8.2.5.6;References;134
8.3;C: Demonstration of Viral Antibodies and Immune Complexes;136
8.3.1;11. Demonstration of Immune Complexes by Capture Enzyme-Linked Immunosorbent Assay;138
8.3.1.1;11.1 Introduction;138
8.3.1.2;11.2 Materials;140
8.3.1.2.1;11.2.1 Reagents;140
8.3.1.2.2;11.2.2 Equipment and Other Consumables;140
8.3.1.2.3;11.2.3 Biological Materials;140
8.3.1.3;11.3 Methods;141
8.3.1.4;11.4 Discussion;141
8.3.1.4.1;11.4.1 Experimental Tips;141
8.3.1.4.2;11.4.2 Precautions;142
8.3.1.4.3;11.4.3 Alternative Materials and/or Methods;142
8.3.1.4.4;11.4.4 Limitations and Future Considerations;142
8.3.1.5;Acknowledgments;142
8.3.1.6;References;143
8.3.2;12. Demonstration of Viral Antibodies by an Immunochromatographic Strip Test;146
8.3.2.1;12.1 Introduction;146
8.3.2.2;12.2 Materials;147
8.3.2.2.1;12.2.1 Reagents;147
8.3.2.2.2;12.2.2 Equipment;148
8.3.2.3;12.3 Methods;148
8.3.2.4;12.4 Discussion;149
8.3.2.5;Acknowledgments;150
8.3.2.6;References;150
8.3.3;13. Rabies Diagnosis: Demonstration of Viral Antibodies by Flow Cytometry;152
8.3.3.1;13.1 Introduction;152
8.3.3.2;13.2 Materials;153
8.3.3.2.1;13.2.1 Reagents;153
8.3.3.2.2;13.2.2 Equipment;154
8.3.3.2.3;13.2.3 Biological Materials;154
8.3.3.3;13.3 Methods;154
8.3.3.3.1;13.3.1 Major Step 1;154
8.3.3.3.2;13.3.2 Major Step 2;154
8.3.3.3.3;13.3.3 Interpretation of Results and Determination of Rabies Virus-Neutralizing Antibodies Levels;155
8.3.3.4;13.4 Discussion;155
8.3.3.4.1;13.4.1 Experimental Tips;157
8.3.3.4.2;13.4.2 Critical Parameters and Troubleshooting;157
8.3.3.4.3;13.4.3 Precautions;157
8.3.3.4.4;13.4.4 Alternative Materials and/or Methods;157
8.3.3.4.5;13.4.5 Time Considerations;158
8.3.3.4.6;13.4.6 Limitations;158
8.3.3.4.7;13.4.7 Future Considerations;158
8.3.3.5;Acknowledgments;158
8.3.3.6;References;159
8.3.4;14. Demonstration of Rabies Antibody by a Latex Agglutination Test;160
8.3.4.1;14.1 Introduction;160
8.3.4.2;14.2 Materials;161
8.3.4.2.1;14.2.1 Reagents;161
8.3.4.2.2;14.2.2 Biological Materials;161
8.3.4.2.3;14.2.3 Equipment;161
8.3.4.3;14.3 Methods;161
8.3.4.3.1;14.3.1 Latex Coating;161
8.3.4.3.2;14.3.2 Slide Agglutination;162
8.3.4.4;14.4 Discussion;162
8.3.4.4.1;14.4.1 Experimental Tips;162
8.3.4.4.2;14.4.2 Technical Notes;163
8.3.4.5;References;164
8.3.5;15. Demonstration of Viral Antibodies by Pseudotype Virus Neutralization;166
8.3.5.1;15.1 Introduction;166
8.3.5.2;15.2 Materials;167
8.3.5.2.1;15.2.1 Reagents;167
8.3.5.2.2;15.2.2 Equipment;168
8.3.5.2.3;15.2.3 Biological Materials;168
8.3.5.3;15.3 Methods;168
8.3.5.3.1;15.3.1 Generation of Pseudotyped Virus Stock;168
8.3.5.3.2;15.3.2 Titration of Pseudotyped Virus Stock;170
8.3.5.3.3;15.3.3 Setting Up a PVNA;170
8.3.5.3.4;15.3.4 Reporter Gene Assay;170
8.3.5.3.5;15.3.5 Interpretation of Results;171
8.3.5.4;15.4 Discussion;172
8.3.5.4.1;15.4.1 Experimental Tips and Critical Parameters;172
8.3.5.4.2;15.4.2 Limitations;173
8.3.5.4.3;15.4.3 Future Considerations;173
8.3.5.5;Acknowledgments;173
8.3.5.6;References;174
8.4;D: Typing/Differentiation of Lyssaviruses;176
8.4.1;16. Sanger Sequencing of Lyssaviruses;178
8.4.1.1;16.1 Background;178
8.4.1.2;16.2 Methodology;180
8.4.1.2.1;16.2.1 Preparing Polymerase Chain Reaction Amplicons for the ABI Genetic Analyzers;180
8.4.1.2.2;16.2.2 Purifying the Polymerase Chain Reaction Product;181
8.4.1.2.3;16.2.3 Big Dye Terminator Cycle Sequencing Reactions;181
8.4.1.2.4;16.2.4 Ethanol Precipitation of Sequencing Reactions;182
8.4.1.2.5;16.2.5 Preparing 96-Well Sample Plate(s) for the Genetic Analyzers;185
8.4.1.2.6;16.2.6 Loading 96-Well Sample Plate(s);185
8.4.1.2.7;16.2.7 Checking Run Reagents;185
8.4.1.2.8;16.2.8 Starting the Run on the 3130xl;186
8.4.1.2.9;16.2.9 Starting the Run on the 3730;186
8.4.1.2.10;16.2.10 Analyzing Output Data;187
8.4.1.3;16.3 Discussion;188
8.4.1.4;Acknowledgments;189
8.4.1.5;References;189
8.4.2;17. Next Generation Sequencing of Lyssaviruses;190
8.4.2.1;17.1 Background;190
8.4.2.2;17.2 Methodology;191
8.4.2.2.1;17.2.1 Viral RNA Extraction;191
8.4.2.2.2;17.2.2 Preparation of cDNA for Sequencing on the Roche 454 FLX(+) Titanium;193
8.4.2.2.3;17.2.3 Analyzing and Annotating 454 Genome Sequences;196
8.4.2.2.4;17.2.4 Depth of Coverage Analysis;200
8.4.2.3;17.3 Discussion;200
8.4.2.4;Acknowledgments;202
8.4.2.5;References;202
8.4.3;18. Genetic Characterization of Rabies Viruses by In Situ Hybridization;204
8.4.3.1;18.1 Introduction;204
8.4.3.2;18.2 Materials;205
8.4.3.3;18.3 Methods;206
8.4.3.3.1;18.3.1 Generation of Control Tissues;206
8.4.3.3.2;18.3.2 Section Preparation;207
8.4.3.3.3;18.3.3 Production and Evaluation of Probes;207
8.4.3.3.4;18.3.4 In Situ Hybridization;207
8.4.3.3.5;18.3.5 Interpretation of Results;208
8.4.3.4;18.4 Discussion;208
8.4.3.4.1;18.4.1 Experimental Tips;208
8.4.3.4.2;18.4.2 Alternative Materials and Methods;208
8.4.3.4.3;18.4.3 Future Considerations;209
8.4.3.5;Acknowledgments;210
8.4.3.6;References;210
8.4.4;19. Oligonucleotide Microarray: Applications for Lyssavirus Speciation;212
8.4.4.1;19.1 Introduction;212
8.4.4.2;19.2 Methodology;214
8.4.4.2.1;19.2.1 Oligonucleotide Design;214
8.4.4.2.2;19.2.2 Microarray Fabrication: In-House Printed Microarray;215
8.4.4.2.3;19.2.3 Microarray Fabrication: In Situ Synthesis (Inkjet Printing) Technology;215
8.4.4.2.4;19.2.4 Nucleic Acid Extraction and DNase Digestion;216
8.4.4.2.5;19.2.5 Reverse Transcription-Polymerase Chain Reaction;217
8.4.4.2.6;19.2.6 Labeling Amplified Target;217
8.4.4.2.7;19.2.7 Microarray Hybridization;218
8.4.4.2.8;19.2.8 Post Hybridization Washes and Image Capture;219
8.4.4.2.9;19.2.9 Data Analysis;219
8.4.4.3;19.3 Discussion;220
8.4.4.4;Acknowledgments;221
8.4.4.5;References;221
8.4.5;20. Demonstration of Lyssavirus Nucleic Acids by Pyrosequencing;224
8.4.5.1;20.1 Introduction;224
8.4.5.2;20.2 Materials;225
8.4.5.2.1;20.2.1 Reagents and Equipment;225
8.4.5.2.2;20.2.2 Diagnostic Samples;225
8.4.5.3;20.3 Methods;225
8.4.5.3.1;20.3.1 Preliminary Phase – Population of the Local Database (IdentiFire Library);225
8.4.5.3.2;20.3.2 Phase 1 – RNA Extraction, OneStep Reverse Transcription-Polymerase Chain Reaction, Gel Electrophoresis Analysis;227
8.4.5.3.3;20.3.3 Phase 2 – Pyrosequencing Reaction and Result Analysis;228
8.4.5.4;20.4 Discussion;229
8.4.5.4.1;20.4.1 Experimental Tips;229
8.4.5.4.2;20.4.2 Critical Parameters and Troubleshooting;229
8.4.5.4.3;20.4.3 Precautions;232
8.4.5.4.4;20.4.4 Alternative Methods;232
8.4.5.4.5;20.4.5 Time Consideration;232
8.4.5.4.6;20.4.6 Limitations;232
8.4.5.4.7;20.4.7 Future Considerations;233
8.4.5.5;Acknowledgments;233
8.4.5.6;References;235
8.5;E: Demonstration of Viral-Induced Changes in Tissues/Organs;238
8.5.1;21. Rabies Diagnosis: MR Imaging;240
8.5.1.1;21.1 Introduction;240
8.5.1.2;21.2 Materials;241
8.5.1.2.1;21.2.1 Magnetic Resonance Imaging Examination;241
8.5.1.2.2;21.2.2 Human and Animal Subjects;241
8.5.1.3;21.3 Methods;242
8.5.1.3.1;21.3.1 Magnetic Resonance Imaging Techniques;242
8.5.1.4;21.4 Discussion;249
8.5.1.5;Acknowledgments;249
8.5.1.6;References;250
9;THREE: RABIES BIOLOGICS;252
9.1;F: Rabies Vaccines for Humans or Other Animals;254
9.1.1;22. Attenuated Vaccines for Veterinary Use;256
9.1.1.1;22.1 Introduction;256
9.1.1.2;22.2 Materials;257
9.1.1.2.1;22.2.1 Reagents;257
9.1.1.2.2;22.2.2 Equipment and Facilities;257
9.1.1.2.3;22.2.3 Biological Materials;257
9.1.1.3;22.3 Methods;259
9.1.1.3.1;22.3.1 Establishment of Master Seed Lot;259
9.1.1.3.2;22.3.2 Upstream Process;259
9.1.1.3.3;22.3.3 Downstream Process;261
9.1.1.3.4;22.3.4 Potency;262
9.1.1.4;22.4 Discussion;262
9.1.1.5;References;263
9.1.2;23. Pox Viral Vectored Vaccines for Rabies;264
9.1.2.1;23.1 Introduction;264
9.1.2.2;23.2 Materials;265
9.1.2.2.1;23.2.1 Reagents;265
9.1.2.2.2;23.2.2 Cells, Viruses, and Other Biological Materials;266
9.1.2.2.3;23.2.3 Equipment;266
9.1.2.3;23.3 Methods;267
9.1.2.3.1;23.3.1 Biological Safety Considerations;267
9.1.2.3.2;23.3.2 Preparation of Vaccinia Virus Transfer Vector Containing a Rabies Virus Glycoprotein Gene;267
9.1.2.3.3;23.3.3 Preparation of Recombinant Vaccinia Viruses;268
9.1.2.3.4;23.3.4 Isolation of Homogenous Recombinant Vaccinia Virus Clones Expressing Rabies Virus Glycoprotein using Mycophenolic Ac ...;269
9.1.2.3.5;23.3.5 Expression Analysis;269
9.1.2.3.6;23.3.6 Culturing of Recombinant Vaccinia Virus for Animal Studies;270
9.1.2.4;23.4 Discussion;270
9.1.2.5;References;271
9.1.3;24. Recombinant Rabies Virus Vaccines;274
9.1.3.1;24.1 Introduction;274
9.1.3.2;24.2 Materials, Methods, and Results;275
9.1.3.2.1;24.2.1 Replication-Competent Recombinant Rabies Virus Vaccines;275
9.1.3.2.2;24.2.2 Replication-Deficient and Replication-Impaired Recombinant Rabies Virus Vaccines;279
9.1.3.3;24.3 Discussion;279
9.1.3.4;References;280
9.1.4;25. Adenoviral Vectors as Vaccine Carriers for Prevention of Rabies;284
9.1.4.1;25.1 Introduction;284
9.1.4.2;25.2 Basic Characteristics of Ad Viruses;285
9.1.4.3;25.3 Types of Ad Vectors;285
9.1.4.4;25.4 Transgene Product for Rabies Vaccines;286
9.1.4.5;25.5 Construction of Ad Vectors;286
9.1.4.6;25.6 Ad Virus Rescue and Expansion;287
9.1.4.7;25.7 Quality Control of Ad Vectors;289
9.1.4.8;25.8 Discussion;291
9.1.4.9;References;292
9.2;G: Rabies Antibodies/Fragments;294
9.2.1;26. Production of Human Monoclonal Antibodies using Mammalian Expression Systems;296
9.2.1.1;26.1 Introduction;297
9.2.1.2;26.2 Materials;297
9.2.1.2.1;26.2.1 Reagents;297
9.2.1.2.2;26.2.2 Equipment;298
9.2.1.2.3;26.2.3 Biological Materials;298
9.2.1.3;26.3 Methods;298
9.2.1.3.1;26.3.1 Cells and Cell Culture;298
9.2.1.3.2;26.3.2 Preparation of Expression Vector;298
9.2.1.3.3;26.3.3 Preparation of Mammalian Cell Stocks;300
9.2.1.3.4;26.3.4 Transfection and Clone Selection;300
9.2.1.3.5;26.3.5 Monoclonal Antibody Expression;301
9.2.1.3.6;26.3.6 Monoclonal Antibody Purification;301
9.2.1.3.7;26.3.7 Monoclonal Antibody Product Testing;303
9.2.1.4;Acknowledgments;307
9.2.1.5;References;308
9.2.2;27. Generation of Immune Globulin Single Variable Domains by Display Technologies;310
9.2.2.1;27.1 Introduction;311
9.2.2.2;27.2 Materials;312
9.2.2.2.1;27.2.1 Reagents;312
9.2.2.2.2;27.2.2 Equipment;312
9.2.2.2.3;27.2.3 Biological Materials;313
9.2.2.2.4;27.2.4 Laboratory Animals;313
9.2.2.3;27.3 Methods;313
9.2.2.3.1;27.3.1 Immunizations;313
9.2.2.3.2;27.3.2 Immune Variable Heavy-Chain Fragment Library Construction for Phage Display;314
9.2.2.3.3;27.3.3 Phage Display for Selection of Rabies Virus-Specific Llama Variable Heavy-Chain Fragment;315
9.2.2.3.4;27.3.4 Periplasmic Expression of Variable Heavy-Chain Fragment;316
9.2.2.4;27.4 Discussion;317
9.2.2.5;27.5 Experimental Tips;317
9.2.2.5.1;27.5.1 Critical Parameters and Troubleshooting;317
9.2.2.5.2;27.5.2 Alternatives;318
9.2.2.5.3;27.5.3 Precautions;318
9.2.2.5.4;27.5.4 Time Considerations;318
9.2.2.5.5;27.5.5 Limitations;318
9.2.2.6;Acknowledgments;319
9.2.2.7;References;320
10;FOUR: APPENDIX;322
10.1;Appendix A;324
10.2;Appendix B;326
10.3;Appendix C;328
10.3.1;C.1 Introduction;328
10.3.2;C.2 Materials;328
10.3.3;C.3 Methods;330
10.3.4;C.4 Discussion;337
10.3.5;Acknowledgments;338
11;Index;340

Leseproben

List of Contributors

Juliano Bordignon,     Department of Virology, Instituto Carlos Chagas, ICC/Fiocruz, Curitiba, Paraná, Brazil; Virology, Instituto Carlos Chagas, ICC/Fiocruz, Curitiba, Paraná, Brazil

Narongsak Chaiyabutr,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand

Florence Cliquet,     French Agency for Food, Environmental and Occupational Health and Safety (Anses), Nancy Laboratory for Rabies and Wildlife, Community Reference Laboratory for Rabies, Community Reference Laboratory for Rabies Serology, OIE Reference Laboratory for Rabies, Technopôle Agricole et Vétérinaire, WHO Collaborating Centre for Research and Management in Zoonoses Control, Malzéville, France

Jessica Coertse,     Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa

Andre Coetzer,     Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, Gauteng, South Africa

Akbar Dastjerdi,     Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom

Cristian De Battisti,     FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (Padova), Italy

Paola De Benedictis,     FAO Reference Centre for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (Padova), Italy

Hans de Haard,     arGEN-X, Zwijnaarde, Belgium

Richard J. Ellis,     Central Sequencing Unit, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom

Hildegund C.J. Ertl,     The Wistar Institute, Philadelphia, PA, USA

Milosz Faber,     Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA

Melina Fischer,     FLI, Institute of Diagnostic Virology, Greifswald-Insel Riems, Germany

Anthony R. Fooks,     University of Liverpool, Department of Clinical Infection, Microbiology and Immunology, Liverpool, United Kingdom; Wildlife Zoonosis and Vector-Borne Disease Research Group, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom; Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom

Conrad M. Freuling,     Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Institute of Molecular Virology and Cell Biology, Greifswald-Insel Riems, Germany

Thiravat Hemachudha,     WHO Collaborating Center for Research and Training on Viral Zoonoses, and Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

Bernd Hoffmann,     FLI, Institute of Diagnostic Virology, Greifswald-Insel Riems, Germany

Anna Hultberg,     arGEN-X, Zwijnaarde, Belgium

Boris Hundt,     IDT Biologika GmbH, Dessau-Rosslau, Germany

Takuya Itou,     Nihon University Veterinary Research Center, Fujisawa, Kanagawa, Japan

Nicholas Johnson,     Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom

Songsri Kasempimolporn,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand

Ivan V. Kuzmin,     Global Alliance for Rabies Control, Manhattan, KS, USA

Jiraporn Laothamatas,     Advanced Diagnostic Imaging Center, and Department of Radiology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

Shampur N. Madhusudana,     Department of Neurovirology, WHO Collaborating Centre for Reference and Research on Rabies, National Institute of Mental Health and Neurosciences, Bangalore, India

Wilfred E. Marissen,     Crucell Holland B.V., Leiden, The Netherlands

Wanda Markotter,     Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, South Africa

Denise A. Marston,     Wildlife Zoonosis and Vector-Borne Disease Research Group, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom

Lorraine M. McElhinney,     Wildlife Zoonosis and Vector-Borne Disease Research Group, Animal Health and Veterinary Laboratories Agency, Addlestone, Surrey, United Kingdom

Kader Muhamuda,     Research Associate, Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

Thomas F. Müller,     Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Molecular Virology and Cell Biology, Greifswald-Insel Riems, Germany

Susan A. Nadin-Davis,     Centre of Expertise for Rabies, Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield, Ottawa, Canada

Thirumeni Nagarajan,     Research and Development, Biological E. Limited, Shameerpet, Hyderabad, Andhra Pradesh, India

Louis H. Nel,     Centre for Emerging and Zoonotic Diseases, National Institute for Communicable Diseases, Sandringham, Gauteng, South Africa; Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, Gauteng, South Africa

Andreas Neubert,     IDT Biologika GmbH, Dessau-Rosslau, Germany

Grit Neumann,     IDT Biologika GmbH, Dessau-Rosslau, Germany

Akira Nishizono,     Department of Microbiology, Faculty of Medicine, Oita University, Japan

Evelyne Picard-Meyer,     French Agency for Food, Environmental and Occupational Health and Safety (Anses), Nancy Laboratory for Rabies and Wildlife, Community Reference Laboratory for Rabies, Community Reference Laboratory for Rabies Serology, OIE Reference Laboratory for Rabies, Technopôle Agricole et Vétérinaire, WHO Collaborating Centre for Research and Management in Zoonoses Control, Malzéville, France

Mani Reeta,     Department of Neurovirology, WHO Collaborating Centre for Reference and Research on Rabies, National Institute of Mental Health and Neurosciences, Bangalore, India

Emmanuelle Robardet,     French Agency for Food, Environmental and Occupational Health and Safety (Anses), Nancy Laboratory for Rabies and Wildlife, Community Reference Laboratory for Rabies, Community Reference Laboratory for Rabies Serology, OIE Reference Laboratory for Rabies, Technopôle Agricole et Vétérinaire, WHO Collaborating Centre for Research and Management in Zoonoses Control, Malzéville, France

Charles Rupprecht,     Ross University School of Veterinary Medicine, Basseterre, St. Kitts, West Indies

Michael Saunders,     arGEN-X, Zwijnaarde, Belgium

Visith Sitprija,     Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand

Catelijne Stortelers,     Ablynx, Zwijnaarde, Belgium National Reference Centre of Rabies, Scientific Institute of Public Helath, Brussels, Belgium

Witaya Sungkarat,     Advanced Diagnostic Imaging Center, and Department of Radiology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

Steven Van Gucht,     arGEN-X, Zwijnaarde, Belgium

Peter Vanlandschoot,     Ablynx, Zwijnaarde, Belgium National Reference Centre of Rabies, Scientific Institute of Public...