Emerging Technologies in Protein and Genomic Material Analysis
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Table of contents
- Cover
- Contentsxiii
- Prefacevii
- List of Contributorsix
- Chapter 1. Enabling Bio-analytical Technologies for Protein and Genomic Material Analysis and their1
- 1.1. Background1
- 1.2. Industrial impact2
- 1.3. Proteomics--the new frontier after the human genome3
- 1.4. Protein chip arrays6
- 1.5. DNA/RNA analysis7
- 1.6. Miniaturized, micro-scale systems8
- 1.7. Concluding remarks9
- 1.8. References10
- Chapter 2. DNA Sequencing: From Capillaries to Microchips11
- 2.1. Introduction11
- 2.2. The early days of automated DNA sequencing12
- 2.3. The advent of capillary gel electrophoresis13
- 2.4. Towards higher throughput: capillary array electrophoresis14
- 2.5. The promise of miniaturization: microchips15
- 2.6. Dealing with the data: bioinformatics17
- 2.7. Conclusions17
- 2.8. References19
- Chapter 3. Phosphoprotein and Phosphoproteome Analysis by Mass Spectrometry21
- 3.1. Introduction21
- 3.2. Phosphoprotein analysis: A challenge for mass spectrometry22
- 3.3. Mass spectrometry based phosphatase/kinase assays25
- 3.4. Phosphoprotein and phosphopeptide enrichment strategies26
- 3.5. Chemical derivatization of phosphopeptides and phosphoproteins27
- 3.6. Selective detection and sequencing of phosphopeptides by tandem mass spectrometry27
- 3.7. Liquid chromatography„tandem mass spectrometry (LC-MS/MS)29
- 3.8. Phosphoproteome analysis by mass spectrometry30
- 3.9. Emerging MS methods tot phosphoprotein analysis31
- 3.10. Relative quantitation of phosphorylation site occupancy by stable isotope labeling of proteins32
- 3.11. Conclusion32
- 3.12. Acknowledgements34
- 3.13. References34
- Chapter 4. Quantitative Peptide Determination Using Column-Switching Capillary Chromatography Interf39
- 4.1. Introduction39
- 4.2. Instrumentation53
- 4.3. Examples of quantitative peptide analysis58
- 4.4. Resume68
- 4.5. Acknowledgements70
- 4.6. References70
- Chapter 5. On-line Continuous-flow Multi-protein Biochemical Assays for the Characterization of Bio-75
- 5.1. Introduction75
- 5.2. Experimental76
- 5.3. Results and discussion79
- 5.4. On-line continuous-flow MS-based biochemical interaction82
- 5.5. Conclusions88
- 5.6. References89
- 5.7. Appendix90
- Chapter 6. Capillary Isoelectric Focusing Developments in Protein Analysis95
- 6.1. Introduction95
- 6.2. Detection96
- 6.3. Micropreparative CIEF103
- 6.4. Internal standards104
- 6.5. Capillary coatings105
- 6.6. CIEF applications106
- 6.7. CIEF-mass spectrometry115
- 6.8. Miniaturized CIEF and chip-IEF122
- 6.9. References130
- Chapter 7. Bio-affinity Extraction for the Analysis of Cytokines and Proteomics Samples135
- 7.1. Introduction to cytokine analysis135
- 7.2. Capillary micro extraction linked to MALDI-TOF MS for the analysis of cytokines at femtomole le136
- 7.3. Application to the analysis of IL-8 and IL-10 in cell samples139
- 7.4. Cytokine analysis conclusions146
- 7.5. Sample preparation for proteomics studies147
- 7.6. Proteomics analysis conclusions150
- 7.7. Acknowledgements152
- 7.8. References152
- Chapter 8. Peptidomics-based Discovery of Endogenous Neuropeptides in the Brain155
- 8.1. Introduction to peptidomics155
- 8.2. Peptidomics sample preparation156
- 8.3. Mass spectrometry of brain tissue peptide extracts157
- 8.4. Tandem mass spectrometry of endogenous neuropeptides158
- 8.5. Simultaneous detection and identification of a large number of endogenous neuropeptides158
- 8.6. Identification of novel neuropeptides158
- 8.7. Identification of 'classical' neuropeptides159
- 8.8. Identification of post-translational modifications of neuropeptides160
- 8.9. Effect of microwave irradiation of brain tissue165
- 8.10. Conclusions166
- 8.11. References166
- Chapter 9. The Beauty of Silicon Micromachined Microstructures Interfaced to MALDI-TOF Mass Spectrom169
- 9.1. Background to miniaturization and microstructure developments within the proteomics research ar169
- 9.2. Introduction to microstructure technology171
- 9.3. Basic concepts of protein determination172
- 9.4. The benefits gained by miniaturization174
- 9.5. Interfacing capillary liquid phase separations to MALDI peptide mass fingerprinting179
- 9.6. Integrated on-line protein workstation182
- 9.7. Integrated microanalytical tool-box components183
- 9.8. Applications to proteomic samples from high sensitivity determinations185
- 9.9. In-vial digestion using nanovial MALDI-target arrays191
- 9.10. Compiled platform developments195
- 9.11. Conclusions195
- 9.12. Future perspectives196
- 9.13. References196
- Chapter 10. Identification and Characterization of Peptides and Proteins Using Fourier Transform Ion199
- 10.1. On DNA and proteins200
- 10.2. On body fluids as sources for clinical markers203
- 10.3. On mass spectrometry206
- 10.4. Experimental approaches212
- 10.5. Proteins in mixtures and human body fluids221
- 10.6. Conclusions233
- 10.7. Acknowledgements235
- 10.8. References235
- Chapter 11. Biological Single Molecule Applications and Advanced Biosensing241
- 11.1. Introduction241
- 11.2. Optical tweezers243
- 11.3. Scanning force spectroscopy248
- 11.4. Advanced bio-sensing using micro-mechanical cantilever arrays254
- 11.5. Conclusion261
- l1.6. Acknowledgements262
- 11.7. References262
- Subject Index265
- Journal of Chromatography Library275
It is widely recognized that analytical technologies and techniques are playing a pioneering role in a range of today's foremost challenging scientific endeavours, including especially biological and biomedical research. Worthy of mention, for example, are the role that high performance separation techniques played in mapping the human genome and the pioneering work done within mass spectrometry.
It is also apparent that state-of-the-art pharmaceutical and biomedical research is the major driving force of the development of new analytical techniques. Advancements in genomics research has provided the opportunity for a call for new drug targets for new technologies, which has speeded up drug discovery and helped to counteract the trend towards inflation of R&D costs.
This book has been designed to be a reference covering a wide range of protein and genomic material analysis techniques. Emerging developments are presented with applications where relevant, and biological examples are included. It was developed to meet the ever growing need for a comprehensive and balanced text on an analytical technique which has generated a tremendous amount of interest in recent years.
In addition, this book also serves as a modern textbook for advanced undergraduate and graduate courses in various disciplines including chemistry, biology and pharmacy.
Authors of the individual chapters are recognized champions of their individual research disciplines and also represent contemporary major research centres in this field.
· Contains state-of-the-art knowledge of the field and detailed descriptions of new technologies
· Provides examples of relevant applications and case studies
· Contributing authors are leading scientists in their own respective research fields
It is also apparent that state-of-the-art pharmaceutical and biomedical research is the major driving force of the development of new analytical techniques. Advancements in genomics research has provided the opportunity for a call for new drug targets for new technologies, which has speeded up drug discovery and helped to counteract the trend towards inflation of R&D costs.
This book has been designed to be a reference covering a wide range of protein and genomic material analysis techniques. Emerging developments are presented with applications where relevant, and biological examples are included. It was developed to meet the ever growing need for a comprehensive and balanced text on an analytical technique which has generated a tremendous amount of interest in recent years.
In addition, this book also serves as a modern textbook for advanced undergraduate and graduate courses in various disciplines including chemistry, biology and pharmacy.
Authors of the individual chapters are recognized champions of their individual research disciplines and also represent contemporary major research centres in this field.
· Contains state-of-the-art knowledge of the field and detailed descriptions of new technologies
· Provides examples of relevant applications and case studies
· Contributing authors are leading scientists in their own respective research fields