Bioconjugate Techniques

Bioconjugate Techniques

3rd Edition - July 25, 2013

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  • Author: Greg Hermanson
  • eBook ISBN: 9780123822406
  • Hardcover ISBN: 9780123822390

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Bioconjugate Techniques, Third Edition, is the essential guide to the modification and cross linking 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.

Key Features

  • Offers a one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab
  • Provides step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates
  • Features full color illustrations
  • Includes a more extensive introduction into the vast field of bioconjugation and one of the most thorough overviews of immobilization chemistry ever presented


Researchers in biochemistry, molecular biology, immunology, pharmacology, cell biology, and microbiology

Table of Contents

  • Dedication

    Preface to the Third Edition


    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



Product details

  • No. of pages: 1200
  • Language: English
  • Copyright: © Academic Press 2013
  • Published: July 25, 2013
  • Imprint: Academic Press
  • eBook ISBN: 9780123822406
  • Hardcover ISBN: 9780123822390

About the Author

Greg Hermanson

Affiliations and Expertise

Director of Technology, Thermo Fisher Scientific, Rockford, IL USA

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