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Biomolecular and Bioanalytical Techniques

Author Vasudevan Ramesh
Release Date: 2019/06/01
ISBN: 9781119483960
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Book Description

An essential guide to biomolecular and bioanalytical techniques and their applications

Biomolecular and Bioanalytical Techniques offers an introduction to, and a basic understanding of, a wide range of biophysical techniques. The text takes an interdisciplinary approach with contributions from a panel of distinguished experts. With a focus on research, the text comprehensively covers a broad selection of topics drawn from contemporary research in the fields of chemistry and biology. Each of the internationally reputed authors has contributed a single chapter on a specific technique. The chapters cover the specific technique’s background, theory, principles, technique, methodology, protocol and applications. 

The text explores the use of a variety of analytical tools to characterise biological samples. The contributors explain how to identify and quantify biochemically important molecules, including small molecules as well as biological macromolecules such as enzymes, antibodies, proteins, peptides and nucleic acids. This book is filled with essential knowledge and explores the skills needed to carry out the research and development roles in academic and industrial laboratories.

  • A technique-focused book that bridges the gap between an introductory text and a book on advanced research methods
  • Provides the necessary background and skills needed to advance the research methods
  • Features a structured approach within each chapter
  • Demonstrates an interdisciplinary approach that serves to develop independent thinking

Written for students in chemistry, biological, medical, pharmaceutical, forensic and biophysical sciences, Biomolecular and Bioanalytical Techniques is an in-depth review of the most current biomolecular and bioanalytical techniques in the field. 

Table of Contents

  1. Cover
  2. List of Contributors
  3. Preface
  4. 1 Principles of Health and Safety and Good Laboratory Practice
    1. 1.1 Introduction
    2. 1.2 Good Laboratory Practice
    3. 1.3 Risk Assessment
    4. 1.4 Chemical Risk Assessment
    5. 1.5 Biological Materials and Genetically Modified Organisms
    6. 1.6 Vacuum Apparatus, Pressure Systems and Associated Glassware
    7. 1.7 Cryogenic Liquefied Gases
    8. 1.8 Compressed Gas Cylinders
    9. 1.9 Electromagnetic Radiation
    10. 1.10 Lasers
    11. 1.11 High Magnetic Fields
    12. 1.12 Sharps
    13. 1.13 Ergonomic Issues
    14. References
  5. 2 Applications of Chemoinformatics in Drug Discovery
    1. 2.1 Significance and Background
    2. 2.2 Computer Representation of Chemical Structures
    3. 2.3 Database Searching
    4. 2.4 Practical Issues on Representation
    5. 2.5 Virtual Screening
    6. 2.6 Ligand‐Based Virtual Screening
    7. 2.7 Protein–Ligand Docking
    8. 2.8 Evaluating Virtual Screening Methods
    9. 2.9 Case Studies of Virtual Screening
    10. 2.10 Conclusions
    11. References
    12. Further Reading
  6. 3 Bioinformatics and Its Applications in Genomics
    1. 3.1 Significance and Short Background
    2. 3.2 Theory/Principles
    3. 3.3 Databases
    4. 3.4 Techniques
    5. 3.5 Applications
    6. 3.6 Concluding Remarks
    7. References
    8. Further Reading
    9. Websites
  7. 4 Gene Cloning for the Analysis of Gene Expression
    1. 4.1 Identifying Target Sequence
    2. 4.2 In Silico Design
    3. 4.3 Primer Design
    4. 4.4 Template Preparation
    5. 4.5 Cloning Methods
    6. 4.6 Uses for Cloned DNA Sequences
    7. 4.7 Verifying Cloned Sequences
    8. 4.8 Applications of Gene Constructs
    9. 4.9 Case Study: Cloning of a Human Missense Variant Exon into a Minigene Splicing Vector
    10. 4.10 Case Study: Epitope Tagging of a Yeast Gene
    11. References
    12. Further Reading
    13. Websites
  8. 5 Proteomic Techniques and Their Applications
    1. 5.1 Significance and Background
    2. 5.2 Principles of Major Proteomics Techniques
    3. 5.3 Methods for Proteomics
    4. 5.4 Applications
    5. 5.5 Concluding Remarks
    6. Acknowledgements
    7. References
    8. Further Reading
    9. Website Resources
  9. 6 Overproduction, Separation and Purification of Affinity‐Tagged Proteins from Escherichia coli
    1. 6.1 Introduction
    2. 6.2 Selecting an Affinity Tag: Glutathione‐S‐Transferase, Maltose‐Binding Protein and Hexa‐Histidine Motifs
    3. 6.3 The pET Vector Series: Archetypal Expression Vectors in E. coli
    4. 6.4 IMAC of a His6‐Tagged Protein: Example Methodology with the ParF DNA Segregation Protein‐Tagged Protein: Example Methodology with the ParF DNA Segregation Protein
    5. 6.5 Production and Purification of a GST‐Tagged Protein: Example Methodology with the C‐Terminal Domain of Yeast RNA Polymerase II
    6. 6.6 Further Purification of Tagged Proteins
    7. 6.7 Alternative Hosts for Protein Production
    8. 6.8 Concluding Remarks
    9. Acknowledgements
    10. References
    11. Further Reading
  10. 7 Chromatography: Separation Techniques in Biology
    1. 7.1 Introduction to Chromatographic Separation
    2. 7.2 General Considerations for Protein Separation by Chromatography
    3. 7.3 Engineering Proteins for Streamlined Chromatographic Separations
    4. 7.4 Example Chromatographic Separations of Biological Samples
    5. 7.5 Other Applications of Chromatography for Biological Sample Preparation and Analysis
    6. References
    7. Further Reading
  11. 8 Synthetic Methodology in Chemical Biology
    1. 8.1 Introduction
    2. 8.2 Peptide Synthesis
    3. 8.3 Amide Bond Synthesis
    4. 8.4 Bioorthogonal Chemistry
    5. 8.5 The Copper‐Catalysed Azide‐Alkyne Cycloaddition Reaction (CuAAC)
    6. 8.6 Unnatural Amino Acid Incorporation
    7. 8.7 Case Studies
    8. 8.8 Conclusion
    9. References
    10. Further Reading
  12. 9 Reaction Chemical Kinetics in Biology
    1. 9.1 Significance
    2. 9.2 Overview of Kinetics and Its Application to Biology
    3. 9.3 Determination of Enzyme Kinetic Mechanisms
    4. 9.4 Technique/Protocol: Determination of Michaelis–Menten Parameters for a Bisubstrate Enzyme and Use of Product Inhibition to Determine Mechanism
    5. 9.5 Case Study: Determination of Michaelis–Menten Parameters for a Bisubstrate Enzyme
    6. 9.6 More Advanced Methods
    7. 9.7 Concluding Remarks
    8. References
  13. 10 Mass Spectrometry and Its Applications
    1. 10.1 Significance
    2. 10.2 Theories and Principles of Biomolecular Mass Spectrometry
    3. 10.3 Techniques and Methodology in Biomolecular Mass Spectrometry
    4. 10.4 Applications
    5. 10.5 Concluding Remarks
    6. References
    7. Further Reading
  14. 11 Applications and Complementarity of Analytical Ultracentrifugation and Light‐Scattering Techniques
    1. 11.1 Introduction
    2. 11.2 Analytical Ultracentrifugation
    3. 11.3 Light Scattering
    4. 11.4 Protocols
    5. 11.5 Applications
    6. 11.6 Conclusions
    7. Acknowledgements
    8. References
    9. Further Reading
  15. 12 Application of Isothermal Titration Calorimetry (ITC) to Biomolecular Interactions
    1. 12.1 Introduction
    2. 12.2 Principles and Theory of ITC
    3. 12.3 Protocols for Design, Implementation and Analysis of ITC Experiments
    4. 12.4 Example Applications of ITC to Analysis of Biomolecular Interactions
    5. 12.5 Concluding Remarks
    6. Acknowledgements
    7. References
    8. Further Reading
    9. Website Resources
  16. 13 An Introduction to Infra‐red and Raman Spectroscopies for Pharmaceutical and Biomedical Studies
    1. 13.1 Significance and Short Background
    2. 13.2 Theory
    3. 13.3 Technique/Methodology/Protocol
    4. 13.4 Applications
    5. 13.5 Concluding Remarks
    6. References
    7. Further Reading
  17. 14 Fluorescence Spectroscopy and Its Applications in Analysing Biomolecular Processes
    1. 14.1 Significance and Background
    2. 14.2 Theory and Principles
    3. 14.3 Techniques, Methodologies and Protocols
    4. 14.4 Case Studies: Fluorescence Spectroscopy to Analyse Membrane Protein Structural Dynamics
    5. 14.5 Concluding Remarks
    6. Acknowledgements
    7. References
    8. Further Reading
  18. 15 Circular Dichroism and Related Spectroscopic Techniques
    1. 15.1 Significance and Background
    2. 15.2 Theory/Principles
    3. 15.3 Technique/Methodology/Protocol
    4. 15.4 Applications
    5. 15.5 Concluding Remarks
    6. References
    7. Further Reading
  19. 16 Principles and Practice in Macromolecular X‐Ray Crystallography
    1. 16.1 Significance and Short Background
    2. 16.2 Theory and Principles: Overview
    3. 16.3 Methodology
    4. 16.4 Applications
    5. 16.5 Concluding Remarks
    6. Acknowledgements
    7. References
    8. Further Reading
  20. 17 Biomolecular NMR Spectroscopy and Structure Determination of DNA
    1. 17.1 Significance and Background
    2. 17.2 Basic NMR Theory
    3. 17.3 Multidimensional NMR Spectroscopy
    4. 17.4 NMR Instrumentation and Experiments [23]
    5. 17.5 Structure and Conformational Parameters of DNA
    6. 17.6 NMR Structure Determination [50–55]
    7. 17.7 Case Study 1: NMR Structure Determination and Conformational Analysis of 17mer Canonical GC DNA
    8. 17.8 Case Study 2: NMR Structure Determination and Conformational Analysis of 13mer 6‐Thioguanine Modified GC DNA [70–74]
    9. 17.9 Conclusion
    10. References
    11. Further Reading
  21. 18 Cryo‐TEM and Biological Structure Determination
    1. 18.1 Significance and Background
    2. 18.2 Theoretical Principles of Biological Cryo‐TEM
    3. 18.3 Experimental Approaches in Biological Cryo‐TEM
    4. 18.4 Cryo‐TEM Case Studies
    5. 18.5 Concluding Remarks
    6. Acknowledgements
    7. References
    8. Website Resources
  22. 19 Computer Modelling and Molecular Dynamics Simulation of Biomolecules
    1. 19.1 Significance
    2. 19.2 Theory and Principles
    3. 19.3 Methodology
    4. 19.4 Applications
    5. 19.5 Concluding Remarks
    6. References
    7. Further Reading
  23. Index
  24. End User License Agreement