Bioconjugate Techniques (eBook)

Greg T. Hermanson (Autor)

eBook Download: PDF

1996 | 1. Auflage
785 Seiten
Elsevier Science (Verlag)
978-0-08-052790-1 (ISBN)

Infos

Bioconjugate Techniques is the essential guide to the modification and crosslinking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Armed with this information and the abundant protocols provided, readers will form unique complexes that can be used for detecting, quantifying, and targeting important analytes.
This book helps readers make: high activity antibody-enzymes conjugates, immunotoxins, immunogen complexes, liposome conjugates, as well as biotinylated molecules, avidin or streptavidin conjugates, colloidal gold labeled proteins, PEG or dextran complexes, labeled oligonucleotide probes, and fluorescently tagged or radiolabeled molecules.

Key Features
* This book is the first to thoroughly capture the entire field of bioconjugate chemistry in a single volume
* Serves as a practical guide to modification and cross-linking technology for research, diagnostics, and therapeutics
* Provides useful, detailed, easy-to-follow, step-by-step protocols
* Contains easy-to-read, and easy-to-understand key concepts for making bioconjugates of all types
* Efficiently covers the chemistry of bioconjugation, the major reagents available for modification and cross-linking, and the application of these reagents to the synthesis of highly active conjugates
* Cites over more than references keyed to concepts covered in the book
* Uses more than 600 figures to illustrate bioconjugate reagents, their reactions, and applications
* Suggests sources for all key reagents

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Bioconjugate Techniques is the essential guide to the modification and crosslinking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Armed with this information and the abundant protocols provided, readers will form unique complexes that can be used for detecting, quantifying, and targeting important analytes. This book helps readers make: high activity antibody-enzymes conjugates, immunotoxins, immunogen complexes, liposome conjugates; as well as biotinylated molecules, avidin or streptavidin conjugates, colloidal gold labeled proteins, PEG or dextran complexes, labeled oligonucleotide probes, and fluorescently tagged or radiolabeled molecules. This book is the first to thoroughly capture the entire field of bioconjugate chemistry in a single volume Serves as a practical guide to modification and cross-linking technology for research, diagnostics, and therapeutics Provides useful, detailed, easy-to-follow, step-by-step protocols Contains easy-to-read, and easy-to-understand key concepts for making bioconjugates of all types Efficiently covers the chemistry of bioconjugation, the major reagents available for modification and cross-linking, and the application of these reagents to the synthesis of highly active conjugates Cites over more than references keyed to concepts covered in the book Uses more than 600 figures to illustrate bioconjugate reagents, their reactions, and applications Suggests sources for all key reagents

Inhalt

Front Cover 1
Bioconjugate Techniques 4
Copyright Page 5
Detailed Contents 10
Preface 22
Acknowledgments 26
Part I: Bioconjugate Chemistry 28
Chapter 1. Functional Targets 30
1. Modification of Amino Acids, Peptides, and Proteins 30
2. Modification of Sugars, Polysaccharides, and Glycoconjugates 54
3. Modification of Nucleic Acids and Oligonucleotides 67
4. Creating Specific Functional Groups 83
5. Blocking Specific Functional Groups 152
Chapter 2. The Chemistry of Reactive Groups 164
1. Amine-Reactive Chemical Reactions 164
2. Thiol-Reactive Chemical Reactions 173
3. Carboxylate-Reactive Chemical Reactions 179
4. Hydroxyl-Reactive Chemical Reactions 181
5. Aldehyde- and Ketone-Reactive Chemical Reactions 186
6. Active Hydrogen-Reactive Chemical Reactions 188
7. Photoreactive Chemical Reactions 190
Part II: Bioconjugate Reagents 194
Chapter 3. Zero-Length Cross-linkers 196
1. Carbodiimides 196
2. Woodward's Reagent K 208
3. N, N'-Carbonyldiimidazole 210
4. Schiff Base Formation and Reductive Amination 212
Chapter 4. Homobifunctional Cross-linkers 214
1. Homobifunctional NHS Esters 215
2. Homobifunctional Imidoesters 230
3. Homobifunctional Sulfhydryl-Reactive Cross-linkers 235
4. Difluorobenzene Derivatives 238
5. Homobifunctional Photoreactive Cross-linkers 241
6. Homobifunctional Aldehydes 241
7. Bis-epoxides 247
8. Homobifuncrional Hydrazides 249
9. Bis-diazonium Derivatives 251
10. Bis-alkylhalides 253
Chapter 5. Heterobifunctional Cross-linkers 255
1. Amine-Reactive and Sulfhydryl-Reactive Cross-linkers 256
2. Carbonyl-Reactive and Sulfhydryl-Reactive Cross-linkers 275
3. Amine-Reactive and Photoreactive Cross-linkers 279
4. Sulfhydryl-Reactive and Photoreactive Cross-linkers 302
5. Carbonyl-Reactive and Photoreactive Cross-linkers 309
6. Carboxylate-Reactive and Photoreactive Cross-linkers 310
7. Arginine-Reactive and Photoreactive Cross-linkers 311
Chapter 6. Trifunctional Cross-linkers 314
1. 4-Azido-2-nitrophenylbiocytin-4-nitrophenyl ester 314
2. Sulfo-SBED 316
Chapter 7. Cleavable Reagent Systems 319
1. Cleavage of Disulfides by Reduction 320
2. Periodate-Cleavable Glycols 321
3. Dithionite-Cleavable Diazo Bonds 322
4. Hydroxylamine-Cleavable Esters 322
5. Base Labile Sulfones 323
Chapter 8. Tags and Probes 324
1. Fluorescent Labels 325
2. Bifunctional Chelating Agents and Radioimmunoconjugates 392
3. Biotinylation Reagents 398
4. lodination Reagents 427
Part III: Bioconjugate Applications 444
Chapter 9. Preparation of Hapten–Carrier Immunogen Conjugates 446
1. The Basis of Immunity 446
2. Types of Immunogen Carriers 448
3. Carbodiimide-Mediated Hapten—Carrier Conjugation 456
4. NHS Ester-Mediated Hapten–Carrier Conjugation 463
5. NHS Ester–Maleimide Heterobifunctional Cross-linker- Mediated Hapten–Carrier Conjugation 466
6. Active-Hydrogen-Mediated Hapten–Carrier Conjugation 473
7. Glutaraldehyde-Mediated Hapten–Carrier Conjugation 480
8. Reductive-Amination-Mediated Hapten–Carrier Conjugation 481
Chapter 10. Antibody Modification and Conjugation 483
1. Preparation of Antibody–Enzyme Conjugates 487
2. Preparation of Labeled Antibodies 514
Chapter 11. Immunotoxin Conjugation Techniques 521
1. Properties and Use of Immunotoxin Conjugates 523
2. Preparation of Immunotoxin Conjugates 514
Chapter 12. Preparation of Liposome Conjugates and Derivatives 555
1. Properties and Use of Liposomes 555
2. Derivatization and Activation of Lipid Components 567
3. Use of Glycolipids and Lectins to Effect Specific Conjugations 575
4. Antigen or Hapten Conjugation to Liposomes 576
5. Preparation of Antibody–Liposome Conjugates 579
6. Preparation of Biotinylated or Avidin-Conjugated Liposomes 617
7. Conjugation of Proteins to Liposomes 583
Chapter 13. Avidin-Biotin Systems 597
1. The Avidin–Biotin Interaction 597
2. Use of Avidin-Biotin in Assay Systems 599
3. Preparation of Avidin or Streptavidin Conjugates 602
4. Preparation of Fluorescently Labeled Avidin or Streptavidin 611
5. Preparation of Hydrazide-Activated Avidin or Streptavidin 616
6. Biotinylation Techniques 617
7. Determination of the Level of Biotinylation 618
Chapter 14. Preparation of Colloidal-Gold-Labeled Proteins 620
1. Properties and Use of Gold Conjugates 620
2. Preparation of Mono-disperse Gold Suspensions for Protein Labeling 624
3. Preparation of Protein A–Gold Complexes 626
4. Preparation of Antibody—Gold Complexes 627
5. Preparation of Lectin–Gold Complexes 628
6. Preparation of Avidin—Gold or Streptavidin–Gold Complexes 629
Chapter 15. Modification with Synthetic Polymers 632
1. Protein Modification with Activated Polyethylene Glycols 633
2. Protein Modification with Activated Dextrans 645
Chapter 16. Enzyme Modification and Conjugation 657
1. Properties of Common Enzymes 657
2. Preparation of Activated Enzymes for Conjugation 662
3. Preparation of Biotinylated Enzymes 664
Chapter 17. Nucleic Acid and Oligonucleotide Modification and Conjugation 666
1. Enzymatic Labeling of DNA 667
2. Chemical Modification of Nucleic Acids and Oligonucleotides 670
References 700
Suppliers of Reagents and Devices for Bioconjugate Applications 754
Index 756

Produkt-Details

• Erscheint lt. Verlag: 15.1.1996
• Sprache: englisch
• Themenwelt: Medizin / Pharmazie ► Medizinische Fachgebiete
• Themenwelt: Studium ► Querschnittsbereiche ► Infektiologie / Immunologie
• Themenwelt: Naturwissenschaften ► Biologie ► Biochemie
• Themenwelt: Naturwissenschaften ► Biologie ► Genetik / Molekularbiologie
• Themenwelt: Naturwissenschaften ► Physik / Astronomie ► Angewandte Physik
• Themenwelt: Technik ► Umwelttechnik / Biotechnologie
• ISBN-10: 0-08-052790-6 / 0080527906
• ISBN-13: 978-0-08-052790-1 / 9780080527901