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Capillary Gel Electrophoresis and Related Microseparation Techniques - 1st Edition - ISBN: 9780444522344

Capillary Gel Electrophoresis and Related Microseparation Techniques

1st Edition

Authors: Andras Guttman László Hajba
Hardcover ISBN: 9780444522344
Imprint: Elsevier Science
Published Date: 1st December 2021
Page Count: 350
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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

  • 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

Academics, industrial analytical chemists, regulatory agencies

Table of Contents

  1. INTRODUCTION
    1.1. History of gel electrophoresis
    1.2. Modes of capillary gel electrophoresis
    1.3. Basic principles and theory of capillary gel electrophoresis
    1.3.1. Electrophoretic Migration
    1.3.2. Secondary Equilibrium
    1.3.3. Efficiency and Resolution
    1.3.4. Capillary gel electrophoresis of DNA
    1.4. Capillary electrophoresis in anticonvective media
    1.5. Band broadening in capillary gel electrophoresis
    1.6. Power dissipation
    1.7. Comparison with slab gel electrophoresis
    1.8. References
    2. SEPARATION MATRIX AND COLUMN TECHNOLOGY
    2.1. Capillary Coatings
    2.1.1. Covalent coatings
    2.1.2. Physical (non-covalent) surface coverage
    2.2. Separation Matrices
    2.2.1. Cross-linked (chemical) gels, structure and properties
    2.2.2. Non cross-linked linear polymers (physical gels)
    2.2.2.1. Non-cross-linked polyacrylamide and agarose
    2.2.2.2. Derivatized celluloses
    2.2.2.3. Polyethylene oxide and polyvinylpyrrolidone
    2.2.3. Copolymers, composite gels and other alternative matrices
    2.3. Techniques for Preparing Gel Filled Capillaries
    2.3.1. Typical gel formulations
    2.3.2. Choosing the appropriate separation media
    2.4. Alternative Matrices (composite gels, pluronics, sol-gel systems, block polymers)
    2.5. References
    3. INSTRUMENTATION
    3.1. Sample Introduction Methods
    3.1.1. Electrokinetic injection into high viscosity gels
    3.1.2. Pressure and vacuum injection into low viscosity polymer solutions
    3.1.3. Sample stacking
    3.1.4. Effect of sample overloading
    3.1.5. Injection related artifacts
    3.2. Detection Systems
    3.2.1. Requirements of capillary gel electrophoresis detectors
    3.2.2. UV absorbance and diode array detection
    3.2.3. Laser induced fluorescence (LIF) detection
    3.2.4. Indirect detection methods
    3.2.5. Electrochemical detection
    3.2.6. Radioisotope detectors and other miscellaneous detection methods
    3.2.7. Coupling to Mass Spectrometers
    3.3. Operation Variables
    3.3.1. Gel concentration, Ferguson plots
    3.3.2. Temperature and voltages, Arrhenius plots
    3.3.3. Capillary dimensions
    3.3.4. Buffer systems
    3.3.5. Organic modifiers, non-aqueous electrophoresis
    3.3.6. Complexing additives
    3.4. References
    4. APPLICATIONS
    4.1. Capillary Gel Electrophoresis of DNA
    4.1.1. Sample purification, injection and separation
    4.1.2. Single stranded oligonucleotides and DNA sequencing
    4.1.3. ds DNA fragments and PCR products
    4.1.4. RNA analysis
    4.1.5. Large chromosomal DNA
    4.1.6. Antisense DNA and phosphodiester oligonucleotides
    4.1.7. Biomedical and forensic applications
    4.2. Capillary Gel Electrophoresis of Proteins
    4.2.1. Fundamentals of capillary SDS gel electrophoresis
    4.2.2. The Ferguson method
    4.2.3. Fluorescent labeling techniques
    4.2.4. Ultrafast separations
    4.2.5. Capillary isoelectric focusing
    4.2.6. Applications in biotechnology
    4.3. Capillary Gel Electrophoresis of Carbohydrates
    4.3.1. Analytical glycobiology
    4.3.2. Release of N and O linked oligosaccharides from glycoproteins
    4.3.3. Fluorophore labeling reactions
    4.3.4. Capillary gel electrophoresis separation of labeled glycans
    4.3.5. Selected applications
    4.4. Capillary Affinity Gel Electrophoresis
    4.4.1. Principles of capillary affinity gel electrophoresis
    4.4.2. Effect of affinity agent concentration and pH
    4.4.3. Chiral additives, types of chiral selectors
    4.4.4. Capillary affinity gel electrophoresis of DNA
    4.4.5. Capillary affinity gel electrophoresis of proteins
    4.4.6. Capillary affinity gel electrophoresis of enantiomers
    4.4.7. Pharmaceutical applications
    4.5. Other, Miscellaneous Applications
    4.6. References
    5. MICROPREPARATIVE CAPILLARY GEL ELECTROPHORESIS
    5.1. Theoretical background of micropreparative applications
    5.2. High-resolution separations
    5.3. Field programming for micropreparative fraction collection
    5.4. Rapid and large scale expressed sequence tag (EST) generation
    5.5. High throughput cloning
    5.6. References
    6. RELATED MICROSEPARATION TECHNIQUES
    6.1. Ultra-Thin-Layer Gel Electrophoresis
    6.1.1. Early miniaturization attempts
    6.1.2. Polyacrylamide gel based systems, DNA sequencing
    6.1.3. Automated ultra-thin-layer gel electrophoresis using agarose gels
    6.1.4. High throughput analysis of dsDNA fragments and proteins
    6.1.5. Two dimensional approaches
    6.1.6. Biomedical applications
    6.2. Gel Electrophoresis in Microfabricated Devices
    6.2.1. Modular system design
    6.2.2. Microfabrication techniques
    6.2.3. Fluid manipulation and injection
    6.2.4. Gel electrophoresis in microchips
    6.2.5. Analysis of single and double stranded DNA
    6.2.6. Analysis of proteins
    6.2.7. Integrated microfabricated biodevices
    6.3. References
    7. APPENDIX. Addresses of some suppliers of instrumentation, chemicals and reagents
    8. GLOSSARY
    9. SUBJECT INDEX

Details

No. of pages:
350
Language:
English
Copyright:
© Elsevier Science 2021
Published:
1st December 2021
Imprint:
Elsevier Science
Hardcover ISBN:
9780444522344

About the Authors

Andras Guttman

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

Horvath Laboratory of Bioseparation Sciences, San Diego, CA, USA

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.

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