Monoclonal antibody and immunosensor technology: the production and application of rodent and human monoclonal antibodies
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| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Amsterdam u.a.
Elsevier
1991
|
| Series: | Laboratory techniques in biochemistry and molecular biology
23 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Literaturverz. S. 401 - 412 |
| Physical Description: | XXV, 427 S. Ill., graph. Darst. |
| ISBN: | 0444814124 0444814132 |
Staff View
MARC
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| 100 | 1 | |a Campbell, Ailsa M. |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Monoclonal antibody and immunosensor technology |b the production and application of rodent and human monoclonal antibodies |c Ailsa M. Campbell |
| 264 | 1 | |a Amsterdam u.a. |b Elsevier |c 1991 | |
| 300 | |a XXV, 427 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a Laboratory techniques in biochemistry and molecular biology |v 23 | |
| 500 | |a Literaturverz. S. 401 - 412 | ||
| 650 | 7 | |a Monoclonal Antibodies |2 cabt | |
| 650 | 7 | |a Techniques |2 cabt | |
| 650 | 4 | |a Anticorps monoclonaux | |
| 650 | 4 | |a Biocapteurs | |
| 650 | 4 | |a Immunologie - Technique | |
| 650 | 4 | |a Antibodies, Monoclonal |v Laboratory Manuals | |
| 650 | 4 | |a Biosensors | |
| 650 | 4 | |a Biotechnology | |
| 650 | 4 | |a Hybridomas |v Laboratory Manuals | |
| 650 | 4 | |a Immunologic Techniques |v Laboratory Manuals | |
| 650 | 4 | |a Monoclonal antibodies | |
| 650 | 0 | 7 | |a Monoklonaler Antikörper |0 (DE-588)4040094-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Biosensor |0 (DE-588)4193016-2 |2 gnd |9 rswk-swf |
| 689 | 0 | 0 | |a Monoklonaler Antikörper |0 (DE-588)4040094-3 |D s |
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| adam_text | IMAGE 1
CONTENTS
PREFACE ACKNOWLEDGEMENTS... ABBREVATIONS
CHAPTER 1. GENERAL PROPERTIES AND APPLICATIONS OF MONOCLONAL ANTIBODIES
I
1.1. HISTORY OF MAB TECHNOLOGY 1
1.2 INTRODUCTION TO MAB TECHNOLOGY 3
1.2.1. THE BASIC METHODOLOGY 3
1.2.2. COMPARISON OF MONOCLONAL ANTIBODIES WITH CONVENTIONAL ANTISERUM 6
1.3 SPECIFICITY/AFFINITY - THE CONTRIBUTION OF MONOCLONAL ANTIBODY 6
1.3.1. THE NATURE OF THE ANTIBODY-ANTIGEN REACTION 6
1.3.2. A SINGLE MONOCLONAL ANTIBODY CAN REACT WITH SEVERAL DISSIMILAR
ANTIGENS 8
1.4. SPECIFICITY/AFFINITY THE CONTRIBUTION OF ANTIGEN 8
1.4.1. AVOIDING MAKING MABS TO SIMILAR ANTIGENS 8
1.4.2. HIGH EPITOPE DENSITY ANTIGENS 10
1.4.3. EXAMPLES OF BAD MABS 11
1.5. SPECIFICITY AFFINITY THE CONTRIBUTION OF THE ASSAY SYSTEM 11
1.6. POOLING OF MABS 12
1.6.1. POOLING OF HIGH-AFFINITY MABS TO GIVE COMPREHENSIVE COVER 12
1.6.2. POOLING OF HIGH-AFFINITY MABS TO FURTHER INCREASE AFFINITY 13
1.6.3. POOLING OF MABS TO PRECIPITATE ANTIGEN 13
1.6.4. POOLING OF LOW-AFFINITY MABS TO ATTEMPT TO GIVE A HIGH-AFFINITY
REAGENT 13
1-7. THE STANDARD NATURE OF MABS 15
1-8. YIELD AND PURITY 15
1-9. HUMAN MONOCLONAL ANTIBODIES (HUMABS) 16
1.9.1. THE REQUIREMENT FOR HUMAN MABS 16
X L LL
IMAGE 2
XIV
1.9.2. BACKGROUND OF HUMAB 1980S DEVELOPMENTS 17
1.9.3. ANTICIPATED FUTURE DEVELOPMENTS 19
1.10. CHIMAERIC BIFUNCTIONAL MONOCLONAL ANTIBODIES 20
1.11. APPLICATIONS OF MABS - DIAGNOSTIC 21
1.11.1. CLINICAL DIAGNOSIS OF SOLUBLE ANTIGENS 21
1.11.2. DIAGNOSIS OF PATHOGENIC BACTERIA, VIRUSES AND PARASITES 23
1.11.3. CELL SURFACE ANTIGENS AND THE CD SYSTEM 23
1.11.4. DIAGNOSTIC APPLICATIONS WITH CELL SURFACE ANTIGENS OTHER THAN
LEUCOCYTE CELL SURFACE ANTIGENS 24
1.11.5. DIAGNOSTICS -PERSONAL KITS 27
1.11.6. DIAGNOSTICS - FORENSIC APPLICATIONS 27
1.11.7. DIAGNOSTICS - IMMUNOSCINTIGRAPHY FOR IN VIVO TUMOUR DIAGNOSIS.
28 1.12. THERAPEUTIC APPLICATIONS 29
1.12.1. INTRODUCTION 29
1.12.2. THERAPY FOR BACTERIAL INFECTION 30
1.12.3. THERAPY FOR VIRAL INFECTION 31
1.12.4. THERAPY FOR TUMOURS 32
1.12.4.1. BACKGROUND 32
1.12.4.2. THERAPY OF LEUKEMIA OR LYMPHOMA 33
1.12.4.3. THERAPY OF SOLID TUMOURS 34
1.12.5. THERAPY FOR PARASITE DISEASES 35
1.13. MABS IN VACCINE DEVELOPMENT STRATEGIES 36
1.13.1. MABS AS REAGENTS TO IDENTIFY CANDIDATE VACCINE MOLECULES 36
1.13.2. MABS ALONE AS VACCINES 37
1.14. MABS AS CATALYSTS (ABZYMES) 39
1.15. MABS IN PURIFICATION 41
1.16. MABS IN BASIC IMMUNOLOGICAL RESEARCH 42
1.17. PATENT CONSIDERATIONS FOR MABS 43
1.17.1. THE PATENT SYSTEM 43
1.17.2. CLASSIC PATENT RULES 44
1.17.3. NOVELTY AND NON-OBVIOUSNESS 45
1.17.4. PRIOR ART 45
1.17.5. HOW SOME MAB PATENTS HAVE OPERATED TO DATE 46
1.17.6. MAB TECHNOLOGY ALREADY FILED FOR PATENT 47
1.17.7. HOW TO PATENT A NEW APPLICATION OF MABS 48
1.18. BUYING A MAB 49
CHAPTER 2. ASSAY TECHNIQUES 51
2. 1. GENERAL ASSAY REQUIREMENTS 51
2.1.1. DEFINITIONS AND CONCEPTS 51
IMAGE 3
XV
2.1.2. TECHNICAL REQUIREMENTS 52
2.2. THEORETICAL CONSIDERATIONS 53
2.2.1. THE NATURE OF THE ANTIBODY-ANTIGEN INTERACTION 53
2.2.2. THE KINETICS OF ANTIBODY-ANTIGEN INTERACTIONS 53
2.2.3. ADDITIONAL CONSIDERATIONS WHICH AFFECT BASIC KINETIC EQUATIONS. .
. 56 2.3. PRACTICAL SCREENING IN THE LAB 59
2.3.1. NUMBERS OF ASSAYS 59
2.3.2. WHEN TO ASSAY 60
2.3.3. CONDITIONS OF ASSAY 60
2.4. ANTIBODY SAMPLING 62
2.5. TYPES OF ASSAY 62
2.6. SOLID-PHASE ASSAYS 63
2.6.1. THE NATURE OF THE SOLID SUPPORT 64
2.6.2. ATTACHMENT OF ANTIGEN 65
2.6.3. BLOCKING OF REMAINING SITES ON THE SOLID SUPPORT 68
2.6.4. STORAGE OF ANTIGEN-BOUND SOLID SUPPORTS 68
2.6.5. INCUBATION WITH ANTIGEN 69
2.6.6. THE NATURE OF THE SECOND ANTIBODY 71
2.6.7. ENZYMES USED IN SCREENING ELISA/DOT-BLOT ASSAYS 73
2.6.8. RADIOACTIVE DETECTION SYSTEMS 80
2.6.9. WASHING 80
2.6.10. DETECTION OF SIGNAL 82
2.7. PROTOCOLS FOR SOLID-PHASE ASSAYS 82
2.7.1. WHETHER TO USE ELISA OR DOT-BLOT 83
2.7.2. ELISA ASSAY FOR IMPURE PROTEIN ANTIGEN 89
2.7.3. ELISA FOR BACTERIAL ANTIGENS 89
2.7.4. ELISA FOR CELLULAR ANTIGENS 90
2.7.5. DOT-BLOTTING FOR PARTICULATE/CELLULAR ANTIGENS 96
2.8. SCREENING FOR CELLULAR ANTIGENS BY FACS 97
2.8.1. SCREENING BY FLOW CYTOMETRY 97
2.8.2. PRESERVATION OF CELLS FOR FACS ASSAY AFTER STAINING 107
2.8.3. PRESERVATION OF CELLS FOR FACS ASSAY BEFORE STAINING 107
2.8.4. DETECTION OF INTRACELLULAR ANTIGENS BY FACS 107
2.8.5. DETECTION OF BACTERIAL ANTIGENS BY FACS 107
2.9. SCREENING FOR CELLULAR ANTIGENS BY IMMUNOCYTOCHEMICAL METHODS 108
2.9.1. PREPARATION OF CELLS AND TISSUES FOR IMMUNOCYTOCHEMISTRY 108
2.9.2. BLOCKING IN IMMUNOHISTOCHEMICAL ASSAYS 110
2.9.3. INCUBATION WITH MAB (OR TEST POLYCLONAL SERUM) 110
2.9.4. CHOICE OF STAINING METHOD I LL
2.9.5. COUNTERSTAINING 112
2.9.6. MOUNTING 113
2.10 SCREENING FOR CATALYTIC ANTIBODIES 113
IMAGE 4
XVI
2.11. SCREENING RECOMBINANT LIBRARIES 114
CHAPTER 3. SELECTION OF ANIMALS, TISSUES AND CELL LINES IN
3.1. ANIMALS IN WHICH HYBRIDOMAS MAY BE GENERATED 117
3.2. WHETHER TO CHOOSE THE RAT OR MOUSE SYSTEM 118
3.2.1. CHOICE OF CELL LINES AS MYELOMA PARTNERS 119
3.2.2. MYELOMA ANTIBODY SYNTHESIS 119
3.2.3. HYBRIDOMA ANTIBODY SECRETION 121
3.2.4. ABILITY OF MYELOMA PARTNERS TO SUPPORT EFFECTIVE FUSION 122
3.2.5. REVERSION FREQUENCY AND STABILITY 122
3.3. SELECTIVE DRUG MARKERS 123
3.3.1. HAT MEDIUM 123
3.3.2. SELECTION FOR AZAGUANINE, THIOGUANINE AND BROMODEOXYURIDINE
RESISTANT MUTANTS 125
3.3.3. SELECTION FOR OUABAIN RESISTANT MUTANTS 126
3.3.4. OTHER METHODS OF SELECTION 126
3.3.5. DOMINANT SELECTION SYSTEMS 126
3.4. THE MOUSE SYSTEM 127
3.4.1. MOUSE CELL LINES TO USE 127
3.4.2. STRAIN OF MOUSE TO USE 129
3.5. THE RAT SYSTEM 130
3.6. THE HUMAN SYSTEM 132
3.6.1. BACKGROUND 132
3.6.2. NEW WAYS TO APPROACH THE PROBLEMS OF HUMAN MAB PRODUCTION. 135
3.6.2.1. HUMAN RODENT CHIMAERAS 135
3.6.2.2. IMMUNISATION 135
3.6.2.3. TISSUE AVAILABILITY 136
3.6.2.4. POLYMERASE CHAIN REACTION 136
3.7. T CELL LINES 137
3.8. TISSUES USED AS SOURCES OF B CELLS 137
CHAPTER 4. IMMUNISATION 139
4.1. HOW THE NATURAL IMMUNE RESPONSE OPERATES 139
4.1.1. EARLY AND IMMEDIATE ANTIBODY RESPONSES 139
4.1.2. AFTER CONTINUED CHALLENGE WITH ANTIGEN 141
4.1.3. TI OR THYMUS INDEPENDENT ANTIGENS 142
4.2. IS IT NECESSARY TO IMMUNISE? 142
IMAGE 5
XV11
4.3. IS IT NECESSARY TO KNOW PRECISELY WHAT THE ANTIGEN IS? 142
4.4. PURITY OF ANTIGEN 144
4.5. THE USE OF PREVIOUSLY DETERMINED SCHEDULES 145
4.6. STRAINS OF RODENT 145
4.7. AGE AND SEX OF RODENT 146
4.8. TOLERANCE, OVERIMMUNISATION AND UNDERIMMUNISATION 147
4.9. ADJUVANTS 148
4.9.1. FREUND S ADJUVANT 148
4.9.2. ALUMINIUM HYDROXIDE (ALUM) 149
4.10. IN VITRO IMMUNISATION 150
4.11. PRODUCTION OF SPECIFIC ANTIBODY ISOTYPES 151
4.11.1. THE REQUIREMENT FOR SPECIFIC ISOTYPES 151
4.11.2. ROUTE OF IMMUNISATION AND TISSUE USED FOR FUSION 151
4.11.3. LYMPHOKINES AND OTHER BIOLOGICAL RESPONSE MODIFIERS 151 4.12.
ROUTES OF IMMUNISATION 152
4.13. SCHEDULES 153
4.14. SPECIFIC EXAMPLES PROTEINS 154
4.14.1. PROTEINS FROM EUKARYOTE SOURCES 154
4.14.2. PROTEINS FROM PROKARYOTE SOURCES 156
4.14.3. PEPTIDES 157
4.15. SPECIFIC EXAMPLES - CARBOHYDRATES 157
4.15.1. SMALL CARBOHYDRATES 157
4.15.2. LARGE CARBOHYDRATES 157
4.16. SPECIFIC EXAMPLES - LIPIDS AND NUCLEIC ACIDS 158
4.17. SPECIFIC EXAMPLES HAPTENS 158
4.17.1. SIZE OF HAPTEN 158
4.17.2. CONJUGATION 159
4.18. SPECIFIC EXAMPLES - WHOLE CELLS AND ORGANISMS 159
4.18.1. VIRUSES 160
4.18.2. BACTERIA 160
4.18.3. WHOLE CELLS 161
4.19. IMMUNISATION OF HUMAN SUBJECTS 162
4.19.1. NON-TOXIC ANTIGENS AND INFECTED INDIVIDUALS 162
4.19.2. TOXIC ANTIGENS 162
CHAPTER 5. CELL CULTURE REQUIREMENTS FOR HYBRIDOMAS 163
5.1. INTRODUCTION 163
5.1.1. SAFETY 163
IMAGE 6
XV111
5.2. BASIC CELL CULTURE REQUIREMENTS 164
5.2.1. PLASTICWARE 165
5.2.2. MEDIA 166
5.2.3. SERA 167
5.2.4. SERUM FREE MEDIA 168
5.2.5. ANTIBIOTICS 169
5.3. BASIC CELL CULTURE TECHNIQUES 170
5.3.1. GENERAL STERILE TECHNIQUE 170
5.3.2. COMPLETE MEDIUM 171
5.3.3. FREEZING AND THAWING OF CELLS 171
5.3.3.1. LIQUID NITROGEN STOCKS 171
5.3.3.2. PREPARATION OF CELLS FOR FREEZING 172
5.3.3.3. THAWING OF CELLS 173
5.3.4. CELL COUNTING AND VIABILITY CHECKS 174
5.4. CONTAMINATION 174
5.4.1. BACTERIAL CONTAMINATION 175
5.4.2. FUNGAL AND YEAST CONTAMINATION 176
5.4.3. CONTAMINATION WITH MYCOPLASMA 176
5.4.3.1. DETECTION OF MYCOPLASMA 178
5.4.3.2. GETTING RID OF MYCOPLASMA - DECONTAMINATION OF EQUIPMENT 179
5.4.3.3. GETTING RID OF MYCOPLASMA - STRATEGIES FOR SAVING A VITAL
HYBRIDOMA 180
5.4.4. CONTAMINATION WITH VIRUSES 181
5.5. FEEDER CELLS 182
5.5.1. FEEDERS FOR MOUSE FUSIONS 182
5.5.2. FEEDERS FOR HUMAN FUSIONS/ TRANSFORMATIONS 187
CHAPTER 6. FUSION PROCEDURES 189
6.1. THE USE OF POLYETHYLENE GLYCOL 189
6.1.1. INTRODUCTION 189
6.1.2. MOLECULAR WEIGHT 189
6.1.3. TEMPERATURE AND PH 190
6.1.4. CONCENTRATION 190
6.1.5. CELL LINES 191
6.1.6. OTHER COMPONENTS OF THE PEG SOLUTION 191
6.2. THE COMPONENTS OF HAT MEDIUM 192
6.3. SELECTION OF HYBRIDOMAS BY FLUORESCENT ACTIVATED CELL SORTING ( F A
C S ) . . .. 193 6.4. PREPARATION OF STOCK SOLUTIONS 194
IMAGE 7
XIX
6.4.1. PREPARATION OF PEG 194
6.4.2. PREPARATION OF HT AND HAT 194
6.4.3. OTHER SELECTIVE MEDIA 195
6.5. FUSION FREQUENCIES AND PLATING DENSITIES 195
6.6. FUSION PROTOCOLS FOR MOUSE EXPERIMENTS 196
6.6.1. PROTOCOL VARIATIONS FOR MOUSE FUSIONS 201
6.6.2. VARIATIONS IN FUSION PROTOCOLS FOR THE RAT SYSTEM 203
6.6.3. VARIATIONS IN FUSION PROTOCOL FOR HUMAN SYSTEMS 203
6.7. ELECTROFUSION 204
CHAPTER 7. ALTERNATIVE STRATEGIES FOR THE IMMORTALISATION OF
ANTIBODY PRODUCING CELLS 205
7.1. INTRODUCTION 205
7.1.1. PURIFICATION OF LYMPHOCYTES FROM COMPLEX TISSUES 206
7.2. PRE-SELECTION OF LYMPHOCYTES WITH DEFINED CELL SURFACE MARKERS OR
ANTIGEN BINDING CAPACITY 207
7.2.1. PRE-SELECTION OF B LYMPHOCYTES FROM T LYMPHOCYTES 209
7.2.1.1. ROSETTING 209
7.2.1.2. PANNING 212
7.2.1.3. NEGATIVE SELECTION USING COMPLEMENT 213
7.2.1.4. FACS SELECTION 214
7.2.1.5. MAGNETIC BEADS 215
7.3. PRE-SELECTION OF T LYMPHOCYTES 215
7.4. TRANSFORMATION OF B LYMPHOCYTES 215
7.4.1. TRANSFORMATION BY EPSTEIN-BARR VIRUS (EBV) 216
7.4.2. SOURCE OF EBV 218
7.4.3. TRANSFORMATION PROCEDURES 219
7.5. POST-TRANSFORMATION STRATEGIES 221
7.5.1. CONTINUOUS CLONING 221
7.5.2. TRANSFORMATION FOLLOWED BY FUSION 222
7.5.3. TRANSFORMATION FOLLOWED BY CLONING OF THE ANTIBODY CODING GENES
223
7.6. TRANSFORMATION WITH ONCOGENES 223
7.7. SHORT TERM CLONING OF B CELLS IN LYMPHOKINE ENRICHED MEDIUM 224
CHAPTER 8. SELECTION AND CLONING 225
8-1. EARLY FEEDING AND ASSAY OF FUSIONS 225
IMAGE 8
XX
8.2. FAILURE OF FUSIONS 228
8.2.1. FAILURE WHERE CLONAL GROWTH IS GOOD 228
8.2.2. FAILURE WITH NO CLONAL GROWTH 229
8.2.3. HOW TO COPE WITH A FUSION WHICH IS TOO SUCCESSFUL 230
8.3. CLONING OF HYBRIDOMAS 231
8.3.1. CLONING BY LIMITING DILUTION 232
8.3.2. CLONING IN SOFT AGAR 234
8.3.3. CLONING BY THE FLUORESCENCE ACTIVATED CELL SORTER (FACS) 235
8.3.4. CLONING OF HUMAN CELLS 235
8.4. FAILURE OF CLONING 235
8.4.1. INTRODUCTION 235
8.4.2. FAILURE WITH NO OR LITTLE GROWTH 236
8.4.3. FAILURE WITH GROWTH 236
8.5. CONTINUATION OF CLONING 236
CHAPTER 9. THE USE OF RECOMBINANT DNA METHODS 239
9.1. BACKGROUND 239
9.2. ANTIBODY GENES 241
9.2.1. ANTIBODY GENES ARE DIFFERENT FROM MOST OTHER GENES IN THAT THEY
UNDERGO A SERIES OF WELL REGULATED REARRANGEMENTS TO CREATE FUNCTIONAL
GENES AS THE B LYMPHOCYTE DIFFERENTIATES 241
9.2.2. ADDITIONAL VARIABILITY IS ADDED DURING ISOTYPE SWITCH 243
9.2.3. TRANSCRIPTION 244
9.2.4. TRANSLATION AND ASSEMBLY 244
9.3. WHETHER TO MAKE WHOLE ANTIBODY, OR ITS SUBFRAGMENTS 245
9.4. THE STEPS INVOLVED IN CLONING OF ANTIBODY GENES 249
9.4.1. THE SOURCE OF THE CDN A OR GENOMIC DNA CODING FOR H AND L CHAINS
249
9.4.2. WHETHER TO CLONE GENOMIC DNA OR CDNA 250
9.4.3. METHODS OF DETECTING ANTIBODY CODING CDNA OR GENOMIC DNA . 250
9.4.4. MANIPULATION OF THE ANTIBODY GENES 251
9.4.5. VECTORS AND SYSTEMS FOR EXPANSION OF THE ANTIBODY CODING GENES
(SHUTTLE VECTORS) 255
9.4.5.1. CHOICE OF VECTOR TYPE 255
9.4.5.2. NECESSARY FUNCTIONS ENCODED BY THE SHUTTLE VECTOR 255 9.4.6.
TRANSFECTION PROCEDURES 257
9.4.7. EXPRESSION VECTORS 258
9.4.7.1. NORMAL ANTIBODY GENE EXPRESSION 258
9.4.7.2. EXPRESSION IN E. COLI AND YEAST 259
IMAGE 9
XXR
9.4.7.3. EXPRESSION IN MYELOMA CELL LINES 259
9.4.7.4. EXPRESSION IN THE BACULOVIRUS SYSTEM 260
9.4.7.5. EXPRESSION IN PLANTS 261
9.5. EXPRESSION IN E. COLI 261
9.5.1. INTRODUCTION 261
9.5.2. SEQUENCES TO BE CLONED 262
9.5.3. VECTORS FOR EXPRESSION IN E. COLI 263
9.6. RECOMBINANT TECHNIQUES INVOLVING THE USE OF PCR 263
9.6.1. INTRODUCTION TO PCR 263
9.6.2. GENERAL LIMITATIONS OF PCR 265
9.6.3. ADDITIONAL PROBLEMS EXPERIENCED WITH THE USE OF PCR AND ANTIBODY
GENES 266
9.6.4. NEW POSSIBILITIES FOR RECOMBINANT ANTIBODY PRODUCTION AS A RESULT
OF PCR 266
9.6.4.1. CLONING THE CDNA OR GENOMIC DNA OF AN ESTABLISHED HYBRIDOMA 266
9.6.4.2. CLONING THE GENES OF AN UNSTABLE HYBRIDOMA 268
9.6.4.3. PCR VARIABLE GENE LIBRARIES 269
9.6.4.4. COMPARISON OF VARIABLE GENE LIBRARIES WITH THE CONVENTIONAL
TECHNIQUE 272
9.7. WHERE RECOMBINANT DNA TECHNOLOGY WILL BE OF VALUE 274
CHAPTER 10. ANTIBODY PRODUCTION AND PURIFICATION 277
10.1. MAINTENANCE OF CELL STOCKS 277
10.2. EXPANSION OF HYBRIDOMAS ON A LABORATORY SCALE (1-20 MG AB) 278
10.2.1. INTRODUCTION 278
10.2.2. EXPANSION IN VITRO FOR PRELIMINARY CHARACTERISATION (1-2 MG) 278
10.3. EXPANSION IN VITRO FOR MORE EXTENSIVE CHARACTERISATION 279
10.4. EXPANSION OF HYBRIDOMAS IN VIVO 280
10.4.1. EXPANSION IN MICE 280
10.4.2. FAILURE OF IN VIVO EXPANSION IN MICE 283
10.4.2.1. FAILURE TO GROW 283
10.4.2.2. GROWTH BUT FAILURE TO SECRETE REQUIRED ANTIBODY. 284 10.4.3.
IN VIVO EXPANSION IN THE RAT 284
10.4.4. EXPANSION OF HUMAN HYBRIDOMAS IN ANIMALS 284
10.5. STORAGE OF MABS 285
10.5.1. STABILITY CHECKS 285
10.5.2. STORAGE AND TRANSPORT OF MABS 286
IMAGE 10
XX11
10.6.
10.5.2.1. GENERAL STORAGE
10.5.2.2. SPECIFIC STORAGE PROBLEMS WITH ASCITES FLUID 10.5.3. TRANSPORT
OF MABS AND HYBRIDOMAS PURIFICATION OF MABS 10.6.1.
10.6.2. 10.6.3.
10.6.4.
10.7.
10.8.
INTRODUCTION MAIN LIKELY CONTAMINANTS CONCENTRATION WITH AMMONIUM
SULPHATE OR CAPRYLIC ACID . MAB CHROMATOGRAPHY
10.6.4.1. MAB PURIFICATION KITS 10.6.4.2. CHROMATOGRAPHY MATRICES
10.6.4.3. FAST PROTEIN LIQUID CHROMATOGRAPHY (FPLC). . 10.6.5.
PURIFICATION WITH DEAE LINKED REAGENTS 10.6.6. PURIFICATION WITH PROTEIN
A OR PROTEIN G 10.6.7. PURIFICATION USING AFFINITY CHROMATOGRAPHY
PURIFICATION OF IGM MABS PURIFICATION OF IGA MABS
286 287 287 288 288 288 289 292 292 292 293 293
295 298 300 301
CHAPTER 11. CHARACTERISATION OF MONOCLONAL ANTIBODIES ...
11.1. 11.2.
11.3.
11.4.
INTRODUCTION DETERMINATION OF ANTIBODY CLASS 11.2.1. CLASS DETERMINATION
BY ELISA 11.2.2. CLASS DETERMINATION BY OUCHTERLONY IMMUNOPRECIPITATION
.
11.2.3. CLASS DETERMINATION BY AGGLUTINATION OF SRBCS 11.2.4. CLASS
DETERMINATION BY ELECTROPHORESIS OF INTERNALLY LABELLED ANTIBODY 11.2.5.
LIGHT CHAIN ANALYSIS PROOF THAT THE ANTIBODY IS MONOCLONAL EPITOPE
ANALYSIS
11.4.1. STRATEGIES BASED ON SYNTHETIC PEPTIDES 11.4.2. DETERMINATION OF
OVERLAPPING EPITOPES 11.4.3. DETERMINATION OF ANTIBODY SPECIFICITY AMONG
A GROUP OF ANTIGENS 11.4.4. ANALYSIS OF EPITOPE SPECIFICITY BY ANTIGEN
MODIFICATION 11.4.5. ANALYSIS OF EPITOPE SPECIFICITY BY WESTERN BLOT
(IMMUNOBLOT) 11.4.5.1. IS BLOTTING AN APPROPRIATE TECHNIQUE FOR MABS?.
11.4.5.2. TYPES OF BLOTTING EQUIPMENT 11.4.5.3. TRANSFER OF PROTEINS
FROM NON-STANDARD GELS . . .
11.4.5.4. MEMBRANES AND TRANSFER BUFFERS 11.4.5.5. BLOCKING AND
DETECTION SYSTEMS
303
303 304 304 305 306
306 310 310 310 311 311
313 316
317 318 319 320
320 321
IMAGE 11
XX111
11.4.6. FAILURE OF BLOTTING 327
11.4.6.1. TECHNICAL FAILURES OF BLOTTING (RARE) 327
11.4.6.2. FUNDAMENTAL DIFFERENCES BETWEEN OPERATOR EXPECTATION AND MAB
RESULT (COMMON) 328
11.5. CHARACTERISATION OF CELL SURFACE ANTIGENS 329
11.6. DETERMINATION OF ANTIBODY AFFINITY 329
11.6.1. QUICK AFFINITY RANKING OF A PANEL 330
11.6.2. METHODS WHICH DO NOT MEASURE AFFINITY BUT MAY APPEAR TO DO SO
331
11.6.3. AFFINITY DETERMINATION FOR MABS DIRECTED AGAINST SOLUBLE
ANTIGENS 331
11.6.3.1. BY IMMUNOPRECIPITATION 331
11.6.3.2. AFFINITY DETERMINATION BY A QUICK ELISA METHOD 333
11.6.3.3. AFFINITY DETERMINATION BY FLUORESCENCE 335
11.6.4. AFFINITY DETERMINATION FOR MABS DIRECTED TO CELLULAR ANTIGENS
335
11.6.4.1. AFFINITY DETERMINATION USING THE FACS 336
11.7. KARYOTYPE ANALYSIS OF HYBRIDOMAS 338
CHAPTER 12. IMMUNOSENSORS 343
12.1. BIOSENSORS 343
12.2. ENZYME BIOSENSORS USING ENZYMES AS SENSING ELEMENTS 344
12.3. THE DIFFERENCE BETWEEN ENZYME BASED SENSING ELEMENTS AND ANTIBODY
BASED SENSING ELEMENTS 345
12.3.1. THE SPECIFICITY OF THE REACTION 345
12.3.2. THE KINETICS OF THE REACTION 345
12.3.3. THE USE OF AN ANTIBODY SENSING CLEMENT WITH AN ENZYME AS PART OF
THE TRANSDUCING SYSTEM 346
12.4. REQUIREMENT AND POTENTIAL APPLICATION OF ANTIBODY BASED
IMMUNOSENSORS 348
12.4.1. POTENTIAL APPLICATIONS 348
12.4.2. IDEAL IMMUNOSENSOR PROPERTIES 349
12.4.3. POSSIBLE DEFECTS IN ANTIBODY BASED IMMUNOSENSORS 350 12.5.
CLASSIFICATION OF IMMUNOSENSOR SYSTEMS 351
12.5.1.1. ELECTROCHEMICAL 351
12.5.1.2. PIEZOELECTRIC 352
12.5.2. GENERAL OPTICAL SYSTEMS 353
12.5.3. INTERNAL REFLECTION SPECTROSCOPY (IRS) OPTICAL SYSTEMS . . ..
353
IMAGE 12
XXIV
12.5.4. CHEMILUMINESCENT/BIOLUMINESCENT SYSTEMS 358
12.5.4.1. HOMOGENEOUS CHEMILUMINESCENT SYSTEMS 359 12.5.4.2.
HETEROGENEOUS CHEMILUMINESCENT SYSTEMS 360 12.6. GENERAL RULES FOR
ADAPTING A MAB OR PANEL OF MABS INTO A BIOSENSOR SYSTEM 361
12.6.1. AFFINITY AND EPITOPE DENSITY 361
12.6.2. THE NATURE OF THE ANTIGEN 361
12.6.3. THE SIZE OF PANEL OF MABS TO BE USED 361
12.6.4. HOMOGENEOUS OR HETEROGENEOUS SYSTEMS 362
12.6.5. ALTERNATIVE DETECTION SYSTEMS 362
12.6.5.1. POLYMERASE CHAIN REACTION (PCR) 362
12.6.5.2. CURRENT SIMPLE DIAGNOSTIC SYSTEMS FOR PROTEINS . 363 12.7.
LABELLING MABS AND THEIR ANTIGENS 364
12.7.1. LABELLING WITH I351 364
12.7.1.1. LABELLING SOLUBLE PROTEINS 364
12.7.1.2. LABELLING CELL MEMBRANES 366
12.7.2. LABELLING WITH ENZYMES 366
12.7.2.1. LABELLING OF HORSERADISH PEROXIDASE (HRP) 366 12.7.2.2.
CROSS-LINKING OF MABS TO ENZYMES BY BIFUNCTIONAL REAGENTS 368
12.7.3. BIOTINYLATION OF MABS 369
12.7.4. LABELLING OF MABS WITH FLUOROCHROMES 370
12.7.5. BINDING MABS TO SOLID SURFACES FOR ASSAY 370
12.7.5.1. BINDING TO BEADS 370
12.7.5.2. BINDING TO GLASS OR QUARTZ 371
APPENDIX 1 - ANIMAL HANDLING TECHNIQUES 373
A.I.I. INTRODUCTION 373
A. 1.2. REQUIREMENT FOR ANIMAL HANDLING LICENCE 373
A.1.3. HANDLING 373
A. 1.3.1. IDENTIFICATION 373
A. 1.3.2. RESTRAINT 374
A. 1.3.3. ANAESTHESIA 374
A. 1.3.4. EUTHANASIA 374
A. 1.4. IMMUNISATION 376
A. 1.4.1. INTRAPERITONEAL 376
A. 1.4.2. SUBCUTANEOUS 377
IMAGE 13
XXV
A. 1.4.3. INTRAVENOUS 378
A. 1.4.4. INTRAMUSCULAR 379
A.L.4.5. INTRADERMAL 379
A.L.4.6. FOOTPAD 379
A. 1.4.7. ORAL 380
A. 1.5. SERUM COLLECTION 381
A.I.5.1. BLEEDING FROM THE TAIL 381
A. 1.5.2. SERUM SEPARATION 382
A. 1.6. REMOVAL OF TISSUES AT SACRIFICE 382
A. 1.6.1. EXSANGUINATION 382
A. 1.6.2. TISSUES CONTAINING IMMUNE CELLS 383
A. 1.7. ASCITES FLUID 387
A.I.7.1. INTRODUCTION 387
A. 1.7.2. TAPPING OF ASCITES FLUID 387
APPENDIX 2 - ADDRESSES OF SUPPLIERS AND MANUFACTURERS 389
APPENDIX 3 - PROTOCOLS FOR POLYACRVLAMIDE GEL ELECTROPHORESIS (PAGE) 393
A.3.1. INTRODUCTION 393
A.3.2. SLAB GEL APPARATUS 393
A.3.3. SDS-PAGE (SODIUM DODECYL SULPHATE-POLYACRYLAMIDE GEL
ELECTROPHORESIS) 394
A.3.4. ISOELECTRIC FOCUSSING OF IMMUNOGLOBULINS 398
A.3.5. ELECTROPHORESIS OF DNA ON AGAROSE 399
A.3.6. AUTORADIOGRAPHY OR FLUOROGRAPHY OF GELS 400
REFERENCES 401
SUBJECT INDEX 413
|
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| author | Campbell, Ailsa M. |
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| author_role | aut |
| author_sort | Campbell, Ailsa M. |
| author_variant | a m c am amc |
| building | Verbundindex |
| bvnumber | BV004737474 |
| callnumber-first | Q - Science |
| callnumber-label | QR186 |
| callnumber-raw | QR186.85 |
| callnumber-search | QR186.85 |
| callnumber-sort | QR 3186.85 |
| callnumber-subject | QR - Microbiology |
| classification_rvk | WC 4150 |
| classification_tum | CHE 810f MED 450f CHE 880f |
| ctrlnum | (OCoLC)28114347 (DE-599)BVBBV004737474 |
| dewey-full | 591.293 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 591 - Specific topics in natural history of animals |
| dewey-raw | 591.293 |
| dewey-search | 591.293 |
| dewey-sort | 3591.293 |
| dewey-tens | 590 - Animals |
| discipline | Biologie Chemie Medizin |
| format | Book |
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| id | DE-604.BV004737474 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T16:16:58Z |
| institution | BVB |
| isbn | 0444814124 0444814132 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-002915187 |
| oclc_num | 28114347 |
| open_access_boolean | |
| owner | DE-19 DE-BY-UBM DE-12 DE-20 DE-91 DE-BY-TUM DE-703 DE-355 DE-BY-UBR DE-188 |
| owner_facet | DE-19 DE-BY-UBM DE-12 DE-20 DE-91 DE-BY-TUM DE-703 DE-355 DE-BY-UBR DE-188 |
| physical | XXV, 427 S. Ill., graph. Darst. |
| publishDate | 1991 |
| publishDateSearch | 1991 |
| publishDateSort | 1991 |
| publisher | Elsevier |
| record_format | marc |
| series | Laboratory techniques in biochemistry and molecular biology |
| series2 | Laboratory techniques in biochemistry and molecular biology |
| spelling | Campbell, Ailsa M. Verfasser aut Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies Ailsa M. Campbell Amsterdam u.a. Elsevier 1991 XXV, 427 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Laboratory techniques in biochemistry and molecular biology 23 Literaturverz. S. 401 - 412 Monoclonal Antibodies cabt Techniques cabt Anticorps monoclonaux Biocapteurs Immunologie - Technique Antibodies, Monoclonal Laboratory Manuals Biosensors Biotechnology Hybridomas Laboratory Manuals Immunologic Techniques Laboratory Manuals Monoclonal antibodies Monoklonaler Antikörper (DE-588)4040094-3 gnd rswk-swf Biosensor (DE-588)4193016-2 gnd rswk-swf Monoklonaler Antikörper (DE-588)4040094-3 s Biosensor (DE-588)4193016-2 s DE-604 Laboratory techniques in biochemistry and molecular biology 23 (DE-604)BV005871299 23 SWB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=002915187&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Campbell, Ailsa M. Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies Laboratory techniques in biochemistry and molecular biology Monoclonal Antibodies cabt Techniques cabt Anticorps monoclonaux Biocapteurs Immunologie - Technique Antibodies, Monoclonal Laboratory Manuals Biosensors Biotechnology Hybridomas Laboratory Manuals Immunologic Techniques Laboratory Manuals Monoclonal antibodies Monoklonaler Antikörper (DE-588)4040094-3 gnd Biosensor (DE-588)4193016-2 gnd |
| subject_GND | (DE-588)4040094-3 (DE-588)4193016-2 |
| title | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies |
| title_auth | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies |
| title_exact_search | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies |
| title_full | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies Ailsa M. Campbell |
| title_fullStr | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies Ailsa M. Campbell |
| title_full_unstemmed | Monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies Ailsa M. Campbell |
| title_short | Monoclonal antibody and immunosensor technology |
| title_sort | monoclonal antibody and immunosensor technology the production and application of rodent and human monoclonal antibodies |
| title_sub | the production and application of rodent and human monoclonal antibodies |
| topic | Monoclonal Antibodies cabt Techniques cabt Anticorps monoclonaux Biocapteurs Immunologie - Technique Antibodies, Monoclonal Laboratory Manuals Biosensors Biotechnology Hybridomas Laboratory Manuals Immunologic Techniques Laboratory Manuals Monoclonal antibodies Monoklonaler Antikörper (DE-588)4040094-3 gnd Biosensor (DE-588)4193016-2 gnd |
| topic_facet | Monoclonal Antibodies Techniques Anticorps monoclonaux Biocapteurs Immunologie - Technique Antibodies, Monoclonal Laboratory Manuals Biosensors Biotechnology Hybridomas Laboratory Manuals Immunologic Techniques Laboratory Manuals Monoclonal antibodies Monoklonaler Antikörper Biosensor |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=002915187&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV005871299 |
| work_keys_str_mv | AT campbellailsam monoclonalantibodyandimmunosensortechnologytheproductionandapplicationofrodentandhumanmonoclonalantibodies |