Elsevier S & T

Bioconjugate Techniques

Hermanson, Greg T.

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Table of contents
  • Copyright Pageiv
  • Contents Overviewvii
  • Detailed Contentsix
  • Prefacexxi
  • Acknowledgmentsxxv
  • Part I: Bioconjugate Chemistry1
  • Chapter 1. Functional Targets3
  • 1. Modification of Amino Acids, Peptides, and Proteins3
  • 2. Modification of Sugars, Polysaccharides, and Glycoconjugates27
  • 3. Modification of Nucleic Acids and Oligonucleotides40
  • 4. Creating Specific Functional Groups56
  • 5. Blocking Specific Functional Groups125
  • Chapter 2. The Chemistry of Reactive Groups137
  • 1. Amine-Reactive Chemical Reactions137
  • 2. Thiol-Reactive Chemical Reactions146
  • 3. Carboxylate-Reactive Chemical Reactions152
  • 4. Hydroxyl-Reactive Chemical Reactions154
  • 5. Aldehyde- and Ketone-Reactive Chemical Reactions159
  • 6. Active Hydrogen-Reactive Chemical Reactions161
  • 7. Photoreactive Chemical Reactions163
  • Part II: Bioconjugate Reagents167
  • Chapter 3. Zero-Length Cross-linkers169
  • 1. Carbodiimides169
  • 2. Woodward's Reagent K181
  • 3. N, N'-Carbonyldiimidazole183
  • 4. Schiff Base Formation and Reductive Amination185
  • Chapter 4. Homobifunctional Cross-linkers187
  • 1. Homobifunctional NHS Esters188
  • 2. Homobifunctional Imidoesters203
  • 3. Homobifunctional Sulfhydryl-Reactive Cross-linkers208
  • 4. Difluorobenzene Derivatives211
  • 5. Homobifunctional Photoreactive Cross-linkers214
  • 6. Homobifunctional Aldehydes214
  • 7. Bis-epoxides220
  • 8. Homobifuncrional Hydrazides222
  • 9. Bis-diazonium Derivatives224
  • 10. Bis-alkylhalides226
  • Chapter 5. Heterobifunctional Cross-linkers228
  • 1. Amine-Reactive and Sulfhydryl-Reactive Cross-linkers229
  • 2. Carbonyl-Reactive and Sulfhydryl-Reactive Cross-linkers248
  • 3. Amine-Reactive and Photoreactive Cross-linkers252
  • 4. Sulfhydryl-Reactive and Photoreactive Cross-linkers275
  • 5. Carbonyl-Reactive and Photoreactive Cross-linkers282
  • 6. Carboxylate-Reactive and Photoreactive Cross-linkers283
  • 7. Arginine-Reactive and Photoreactive Cross-linkers284
  • Chapter 6. Trifunctional Cross-linkers287
  • 1. 4-Azido-2-nitrophenylbiocytin-4-nitrophenyl ester287
  • 2. Sulfo-SBED289
  • Chapter 7. Cleavable Reagent Systems292
  • 1. Cleavage of Disulfides by Reduction293
  • 2. Periodate-Cleavable Glycols294
  • 3. Dithionite-Cleavable Diazo Bonds295
  • 4. Hydroxylamine-Cleavable Esters295
  • 5. Base Labile Sulfones296
  • Chapter 8. Tags and Probes297
  • 1. Fluorescent Labels298
  • 2. Bifunctional Chelating Agents and Radioimmunoconjugates365
  • 3. Biotinylation Reagents371
  • 4. lodination Reagents400
  • Part III: Bioconjugate Applications417
  • Chapter 9. Preparation of Hapten–Carrier Immunogen Conjugates419
  • 1. The Basis of Immunity419
  • 2. Types of Immunogen Carriers421
  • 3. Carbodiimide-Mediated Hapten„Carrier Conjugation429
  • 4. NHS Ester-Mediated Hapten–Carrier Conjugation436
  • 5. NHS Ester–Maleimide Heterobifunctional Cross-linker- Mediated Hapten–Carrier Conjugation439
  • 6. Active-Hydrogen-Mediated Hapten–Carrier Conjugation446
  • 7. Glutaraldehyde-Mediated Hapten–Carrier Conjugation453
  • 8. Reductive-Amination-Mediated Hapten–Carrier Conjugation454
  • Chapter 10. Antibody Modification and Conjugation456
  • 1. Preparation of Antibody–Enzyme Conjugates460
  • 2. Preparation of Labeled Antibodies487
  • Chapter 11. Immunotoxin Conjugation Techniques494
  • 1. Properties and Use of Immunotoxin Conjugates496
  • 2. Preparation of Immunotoxin Conjugates487
  • Chapter 12. Preparation of Liposome Conjugates and Derivatives528
  • 1. Properties and Use of Liposomes528
  • 2. Derivatization and Activation of Lipid Components540
  • 3. Use of Glycolipids and Lectins to Effect Specific Conjugations548
  • 4. Antigen or Hapten Conjugation to Liposomes549
  • 5. Preparation of Antibody–Liposome Conjugates552
  • 6. Preparation of Biotinylated or Avidin-Conjugated Liposomes590
  • 7. Conjugation of Proteins to Liposomes556
  • Chapter 13. Avidin-Biotin Systems570
  • 1. The Avidin–Biotin Interaction570
  • 2. Use of Avidin-Biotin in Assay Systems572
  • 3. Preparation of Avidin or Streptavidin Conjugates575
  • 4. Preparation of Fluorescently Labeled Avidin or Streptavidin584
  • 5. Preparation of Hydrazide-Activated Avidin or Streptavidin589
  • 6. Biotinylation Techniques590
  • 7. Determination of the Level of Biotinylation591
  • Chapter 14. Preparation of Colloidal-Gold-Labeled Proteins593
  • 1. Properties and Use of Gold Conjugates593
  • 2. Preparation of Mono-disperse Gold Suspensions for Protein Labeling597
  • 3. Preparation of Protein A–Gold Complexes599
  • 4. Preparation of Antibody„Gold Complexes600
  • 5. Preparation of Lectin–Gold Complexes601
  • 6. Preparation of Avidin„Gold or Streptavidin…Gold Complexes602
  • Chapter 15. Modification with Synthetic Polymers605
  • 1. Protein Modification with Activated Polyethylene Glycols606
  • 2. Protein Modification with Activated Dextrans618
  • Chapter 16. Enzyme Modification and Conjugation630
  • 1. Properties of Common Enzymes630
  • 2. Preparation of Activated Enzymes for Conjugation635
  • 3. Preparation of Biotinylated Enzymes637
  • Chapter 17. Nucleic Acid and Oligonucleotide Modification and Conjugation639
  • 1. Enzymatic Labeling of DNA640
  • 2. Chemical Modification of Nucleic Acids and Oligonucleotides643
  • References673
  • Suppliers of Reagents and Devices for Bioconjugate Applications727
  • Index729
Book details
  • Vendor Elsevier S & T
  • SKU 9780123423351
  • ISBN-13 9780080527901
  • Author Hermanson, Greg T.
  • Category Science
  • Subject Research & Methodology

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Description eBook delivery & access Reviews
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