Capillary Gel Electrophoresis
- 1st Edition - December 4, 2021
- Latest edition
- Authors: Andras Guttman, László Hajba
- Language: English
Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent… Read more
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Description
Description
Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses.
Key features
Key features
- Covers all theoretical and practical aspects of capillary gel electrophoresis
- Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods
- Teaches readers how to use the technology and select methods that are ideal for fundamental problems
- Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
Readership
Readership
Academics, industrial analytical chemists, regulatory agencies
Table of contents
Table of contents
Preface
1. Introduction
1.1 Short history of capillary gel electrophoresis
1.2 The capillary gel electrophoresis system
1.2.1 Basic setup
1.2.2 The separation capillary
1.2.3 Power supply
1.2.4 Injection
1.2.5 Detection
1.2.6 Data acquisition
1.3 Separation modes in gel-filled capillary columns
1.3.1 CGE for molecular sieving
1.3.2 Capillary gel isoelectric focusing
1.3.3 Capillary gel isotachophoresis
1.3.4 Capillary affinity gel electrophoresis
1.4 Comparison with slab gel electrophoresis
References
2. Basic principles of capillary gel electrophoresis
2.1 The electrophoretic migration
2.2 Efficiency and resolution
2.3 Band broadening in capillary gel electrophoresis
2.4 Temperature effects and power dissipation
2.5 Complexation equilibrium
2.6 Electric field-mediated migration of biopolymers in sieving matrices
2.7 Micropreparative capillary gel electrophoresis
References
3. Separation matrix and column technology
3.1 Gels and polymer networks
3.1.1 Cross-linked (chemical) gels
3.1.2 Noncross-linked polymer solutions (physical gels)
3.2 Alternative matrices (composite gels, pluronics, agarose,
sol
3.2.1 Agarose-based gel compositions
3.2.2 Other alternative matrices
3.3 Capillary coatings
3.3.1 Covalent (chemical) coatings
3.3.2 Physical (noncovalent) surface coverage
3.4 Techniques for preparing gel-filled capillaries
References
4. Instrumentation
4.1 Sample introduction methods
4.1.1 Electrokinetic injection into high-viscosity gels
4.1.2 Hydrodynamic injection into low-viscosity polymer solutions
4.1.3 Sample preconcentration
4.1.4 Effect of sample overloading
4.1.5 Injection-related artifacts
4.2 Detection systems
4.2.1 UV absorbance and diode array detection
4.2.2 Laser-induced fluorescence detection
4.2.3 Indirect detection methods
4.2.4 Electrochemical and conductivity detection
4.2.5 Miscellaneous detector systems
4.2.6 Coupling to mass spectrometers
4.3 Operation variables
4.3.1 Gel concentration
4.3.2 Separation voltage and temperature
4.3.3 Capillary dimensions
4.3.4 Buffer systems
4.3.5 Nonaqueous electrophoresis, organic modifiers
References
5. Applications
5.1 Capillary gel electrophoresis of DNA
5.1.1 Analysis of single-stranded oligonucleotides
5.1.2 dsDNA fragments and PCR products
5.1.3 Large chromosomal DNA and pulsed-field electrophoresis
5.1.4 Antisense DNA and modified oligonucleotides
5.1.5 Biomedical and forensic applications
5.2 Capillary gel electrophoresis of proteins
5.2.1 Capillary SDS gel electrophoresis
5.2.2 Ultrafast separations
5.2.3 Capillary gel isoelectric focusing of proteins
5.2.4 Fluorescent labeling of proteins for SDS-CGE
5.2.5 Applications in biotechnology
5.3 Capillary gel electrophoresis of carbohydrates
5.3.1 Analytical glycobiology
5.3.2 Release of N- and O-linked oligosaccharides from glycoproteins
5.3.3 Fluorophore labeling of carbohydrates
5.3.4 Capillary gel electrophoresis separation of oligosaccharides
5.3.5 Selected other applications
5.4 Capillary affinity gel electrophoresis
5.4.1 Chiral affinity additives and chiral selectors
5.4.2 Capillary affinity gel electrophoresis of DNA
5.4.3 Other capillary affinity gel electrophoresis applications
References
6. Related microseparation techniques
6.1 Ultrathin-layer gel electrophoresis
6.1.1 DNA analysis
6.1.2 Protein analysis
6.2 Multidimensional approaches
References
7. Appendix
Manufacturers’ directory
Further reading
Abbreviations
Index
1. Introduction
1.1 Short history of capillary gel electrophoresis
1.2 The capillary gel electrophoresis system
1.2.1 Basic setup
1.2.2 The separation capillary
1.2.3 Power supply
1.2.4 Injection
1.2.5 Detection
1.2.6 Data acquisition
1.3 Separation modes in gel-filled capillary columns
1.3.1 CGE for molecular sieving
1.3.2 Capillary gel isoelectric focusing
1.3.3 Capillary gel isotachophoresis
1.3.4 Capillary affinity gel electrophoresis
1.4 Comparison with slab gel electrophoresis
References
2. Basic principles of capillary gel electrophoresis
2.1 The electrophoretic migration
2.2 Efficiency and resolution
2.3 Band broadening in capillary gel electrophoresis
2.4 Temperature effects and power dissipation
2.5 Complexation equilibrium
2.6 Electric field-mediated migration of biopolymers in sieving matrices
2.7 Micropreparative capillary gel electrophoresis
References
3. Separation matrix and column technology
3.1 Gels and polymer networks
3.1.1 Cross-linked (chemical) gels
3.1.2 Noncross-linked polymer solutions (physical gels)
3.2 Alternative matrices (composite gels, pluronics, agarose,
sol
3.2.1 Agarose-based gel compositions
3.2.2 Other alternative matrices
3.3 Capillary coatings
3.3.1 Covalent (chemical) coatings
3.3.2 Physical (noncovalent) surface coverage
3.4 Techniques for preparing gel-filled capillaries
References
4. Instrumentation
4.1 Sample introduction methods
4.1.1 Electrokinetic injection into high-viscosity gels
4.1.2 Hydrodynamic injection into low-viscosity polymer solutions
4.1.3 Sample preconcentration
4.1.4 Effect of sample overloading
4.1.5 Injection-related artifacts
4.2 Detection systems
4.2.1 UV absorbance and diode array detection
4.2.2 Laser-induced fluorescence detection
4.2.3 Indirect detection methods
4.2.4 Electrochemical and conductivity detection
4.2.5 Miscellaneous detector systems
4.2.6 Coupling to mass spectrometers
4.3 Operation variables
4.3.1 Gel concentration
4.3.2 Separation voltage and temperature
4.3.3 Capillary dimensions
4.3.4 Buffer systems
4.3.5 Nonaqueous electrophoresis, organic modifiers
References
5. Applications
5.1 Capillary gel electrophoresis of DNA
5.1.1 Analysis of single-stranded oligonucleotides
5.1.2 dsDNA fragments and PCR products
5.1.3 Large chromosomal DNA and pulsed-field electrophoresis
5.1.4 Antisense DNA and modified oligonucleotides
5.1.5 Biomedical and forensic applications
5.2 Capillary gel electrophoresis of proteins
5.2.1 Capillary SDS gel electrophoresis
5.2.2 Ultrafast separations
5.2.3 Capillary gel isoelectric focusing of proteins
5.2.4 Fluorescent labeling of proteins for SDS-CGE
5.2.5 Applications in biotechnology
5.3 Capillary gel electrophoresis of carbohydrates
5.3.1 Analytical glycobiology
5.3.2 Release of N- and O-linked oligosaccharides from glycoproteins
5.3.3 Fluorophore labeling of carbohydrates
5.3.4 Capillary gel electrophoresis separation of oligosaccharides
5.3.5 Selected other applications
5.4 Capillary affinity gel electrophoresis
5.4.1 Chiral affinity additives and chiral selectors
5.4.2 Capillary affinity gel electrophoresis of DNA
5.4.3 Other capillary affinity gel electrophoresis applications
References
6. Related microseparation techniques
6.1 Ultrathin-layer gel electrophoresis
6.1.1 DNA analysis
6.1.2 Protein analysis
6.2 Multidimensional approaches
References
7. Appendix
Manufacturers’ directory
Further reading
Abbreviations
Index
Product details
Product details
- Edition: 1
- Latest edition
- Published: December 8, 2021
- Language: English
About the authors
About the authors
AG
Andras Guttman
András Guttman, MTA-Lendulet Professor of Translational Glycomics at the Research Institute for Biomolecular and Chemical Engineering at University of Pannonia (Veszprem, Hungary), also heading the Horváth Csaba Memorial Laboratory of Bioseparation Sciences. His work is focused on capillary gel electrophoresis for glycomics and glycoproteomics analysis of samples of biomedical and biopharmaceutical interests. Dr Guttman has over 350 scientific publications, wrote 35 book chapters, edited 4 textbooks and holds 25 patents. He is an external member of the Hungarian Academy of Sciences, Editor in Chief for Current Molecular Medicine, serves as editorial board member for a dozen scientific journals and on the board of several international scientific organizations. He has been recognized by numerous awards including the Analytical Chemistry Award of the Hungarian Chemical Society, named as Fulbright Scholar, received the CASSS CE Pharm Award, the Arany Janos medal of the Hungarian Academy of Sciences, the Pro Scientia award of the University of Pannonia and the Dennis Gabor Award of the Novofer Foundation. Dr Guttman is also the recipient of the Dal Nogare Award of the Delaware Valley Chromatography Forum and the Grand prize of the Swedish Chamber of Commerce.
Affiliations and expertise
Horváth Csaba Memorial Laboratory of Bioseparation Sciences Research Center for Molecular Medicine, Faculty of Medicine University of Debrecen
98 Nagyerdei Krt, H-4032, Debrecen, HungaryLH
László Hajba
László Hajba holds a PhD in analytical and environmental chemistry from University of Pannonia, awarded in 2008, his research topic was the vibrational spectroscopic, theoretical and structural study of organometallic molecules. Later he carried out research in the field of biospectroscopy and chemometrics at the same university. He performed infrared spectroscopic investigation of human hair and skin, identification of cancer specific spectroscopic markers with chemometric methods. In 2010 he joined the biotechnology research group at Research Institute of Chemical and Process Engineering, University of Pannonia. In 2013 he started his work in the field of microfluidics and bioseparations at the Translational Glycomics Research Group. He and his coworkers developed a fully automated linear polyacrylamide capillary coating method for high efficiency capillary electrophoretic separation of cell surface proteins and the glycosimilarity index for the similarity measure of the glycan structures of therapeutic proteins. He received the Outstanding Scientist Award from the Hungarian Academy of Sciences in 2019. He has 47 peer reviewed scientific publication with more than 650 independent citations.
Affiliations and expertise
Translational Glycomics Research Group Research Institute of Biomolecular and Chemical Engineering, University of Pannonia 10 Egyetem Street, H-8200 Veszprém, HungaryView book on ScienceDirect
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