Bioconjugate Techniques

Dedication

Preface to the Third Edition

Acknowledgments

Important Information

Health and Safety

Intellectual Property

Chapter 1. Introduction to Bioconjugation

1 What is Bioconjugation?

2 Bioconjugation Strategy and Design

3 The Applications of Bioconjugates

4 Summary

Chapter 2. Functional Targets for Bioconjugation

1 Modification of Amino Acids, Peptides, and Proteins

2 Modification of Sugars, Polysaccharides, and Glycoconjugates

3 Modification of Nucleic Acids and Oligonucleotides

4 Creating Specific Functionalities

5 Blocking or Protecting Groups

Chapter 3. The Reactions of Bioconjugation

1 Amine Reactions

2 Thiol Reactions

3 Carboxylate Reactions

4 Hydroxyl Reactions

5 Aldehyde and Ketone Reactions

6 Active Hydrogen Reactions

7 PhotoChemical Reactions

8 Cycloaddition Reactions

Chapter 4. Zero-Length Crosslinkers

1 Carbodiimides

2 Woodward’s Reagent K

3 N, N′-Carbonyl Diimidazole

4 Schiff Base Formation and Reductive Amination

Chapter 5. Homobifunctional Crosslinkers

1 Homobifunctional NHS Esters

2 Homobifunctional Imidoesters

3 Homobifunctional Sulfhydryl Reactive Crosslinkers

4 Difluorobenzene Derivatives

5 Homobifunctional Photoreactive Crosslinkers

6 Homobifunctional Aldehydes

7 BIS-Epoxides

8 Homobifunctional Hydrazides

9 BIS-Diazonium Derivatives

10 BIS-Alkylhalides

Chapter 6. Heterobifunctional Crosslinkers

1 Amine-Reactive and Sulfhydryl-Reactive Crosslinkers

2 Carbonyl Reactive and Sulfhydryl Reactive Crosslinkers

3 Amine-Reactive and Photoreactive Crosslinkers

4 Sulfhydryl-Reactive and Photoreactive Crosslinkers

5 Carbonyl-Reactive and Photoreactive Crosslinkers

6 Carboxylate-Reactive and Photoreactive Crosslinkers

7 Arginine-Reactive and Photoreactive Crosslinkers

Chapter 7. Trifunctional Crosslinkers

1 4-Azido-2-Nitrophenylbiocytin-4-Nitrophenyl Ester

2 Sulfo-SBED

3 MTS–ATF–Biotin and MTS–ATF–LC-Biotin

4 Hydroxymethyl Phosphine Derivatives

5 Tricepts Reagent

Chapter 8. Dendrimers and Dendrons

1 Dendrimer Construction

2 Conjugation to Dendrimers

3 Dendrimer–Chelate Derivatives for Imaging Applications

4 Dendrimer Derivatives as Surface Modification Agents

5 Dendrimer Fluorescent Quantum Dots

Chapter 9. Cross-Bridges and Cleavable Reagent Systems

1 Cleavage of Disulfides by Reduction

2 Periodate-Cleavable Glycols

3 Dithionite-Cleavable Bonds

4 Hydroxylamine-Cleavable Esters

5 Base Labile Sulfones

6 Acyl Hydrazone-Cleavable Linkers

7 Photocleavable Linkers

Chapter 10. Fluorescent Probes

1 Fluorescein Derivatives

2 Rhodamine Derivatives

3 Coumarin Derivatives

4 Boron Dipyrromethene Derivatives

5 Pyrene Derivatives

6 Naphthalimide Derivatives

7 Phycobiliprotein Derivatives

8 Cyanine Dye Derivatives

9 Lanthanide Chelates for Time-Resolved Fluorescence

10 Quantum Dot Nanocrystals

Chapter 11. (Strept)avidin–Biotin Systems

1 The (Strept)Avidin–Biotin Interaction

2 Use of (Strept)Avidin–Biotin Interactions in Assay Systems

3 Preparation of (Strept)Avidin Conjugates

4 Preparation of Fluorescently Labeled (Strept)Avidin

5 Preparation of Hydrazide Activated (Strept)Avidin

6 Biotinylation Techniques

Chapter 12. Isotopic Labeling Techniques

1 Bifunctional Chelating Agents and Radioimmunoconjugates

2 Iodination Reagents

3 Mass Tags and Isotope Tags

Chapter 13. Silane Coupling Agents

1 Silane Reaction Strategies

2 Functional Silane Compounds

Chapter 14. Microparticles and Nanoparticles

1 Particle Types

2 Particle Characteristics and Stability

3 Particle Concentration

4 Polymeric Microspheres and Nanospheres

5 Silica Particles

6 Gold Particles

Chapter 15. Immobilization of Ligands on Chromatography Supports

1 Support Materials Used for Affinity Chromatography

2 Activation and Coupling of Affinity Ligands to Chromatography Supports

Chapter 16. Buckyballs, Fullerenes, and Carbon Nanotubes

1 Buckyballs and Fullerenes

2 Carbon Nanotubes

Chapter 17. Chemoselective Ligation; Bioorthogonal Reagents

1 Diels–Alder Reagent Pairs

2 Hydrazine–Aldehyde Reagent Pairs

3 Aminooxy–Aldehyde Reagent Pairs

4 Boronic Acid–Salicylhydroxamate Reagent Pairs

5 Click Chemistry: Cu(I)- Promoted Azide–Alkyne [3+2] Cycloaddition

6 Staudinger Ligation

7 Native Chemical Ligation

Chapter 18. PEGylation and Synthetic Polymer Modification

1 Discrete PEG Reagents

2 Polymer Modification Reagents

Chapter 19. Vaccines and Immunogen Conjugates

1 The Basis of Immunity

2 Types of Immunogen Carriers

3 Carbodiimide-Mediated Hapten–Carrier Conjugation

4 NHS Ester-Mediated Hapten–Carrier Conjugation

5 NHS Ester–Maleimide Heterobifunctional Crosslinker-Mediated Hapten–Carrier Conjugation

6 Active-Hydrogen-Mediated Hapten–Carrier Conjugation

7 Glutaraldehyde-Mediated Hapten–Carrier Conjugation

8 Reductive Amination-Mediated Hapten–Carrier Conjugation

Chapter 20. Antibody Modification and Conjugation

1 Preparation of Antibody–Enzyme Conjugates

2 Preparation of Labeled Antibodies

3 Immunotoxin Conjugates

Chapter 21. Liposome Conjugates and Derivatives

1 Properties and Use of Liposomes

2 Derivatization and Activation of Lipid Components

3 Use of Glycolipids and Lectins to Effect Specific Conjugations

4 Antigen or Hapten Conjugation to Liposomes

5 Preparation of Antibody–Liposome Conjugates

6 Preparation of Biotinylated or (strept)avidin-Conjugated Liposomes

7 Conjugation of Proteins to Liposomes

Chapter 22. Enzyme Modification and Conjugation

1 Properties of Common Enzymes

2 Preparation of Activated Enzymes for Conjugation

3 Preparation of Biotinylated Enzymes

Chapter23. Nucleic Acid and Oligonucleotide Modification and Conjugation

1 Enzymatic Labeling of DNA

2 Chemical Modification of Nucleic Acids and Oligonucleotides

Chapter 24. Bioconjugation in the Study of Protein Interactions

1 Homobifunctional Crosslinking Agents

2 Use of Photoreactive Crosslinkers to Study Protein Interactions

3 Trifunctional Label Transfer Reagents

4 Metal Chelates in the Study of Protein Interactions

References

Index