Capillary Gel Electrophoresis and Related Microseparation Techniques - 1st Edition - ISBN: 9780444522344, 9780080931357

Capillary Gel Electrophoresis and Related Microseparation Techniques

1st Edition

Authors: Andras Guttman
eBook ISBN: 9780080931357
Hardcover ISBN: 9780444522344
Imprint: Elsevier Science
Published Date: 1st February 2020
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, as well as related microseparation methods. It gives readers a better understanding of how to utilize this technology, and determine which method will provide the best technical solutions to their 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, as well as related microseparation methods
  • Teaches readers how to use the technology, and how to select methods 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 2020
Published:
Imprint:
Elsevier Science
eBook ISBN:
9780080931357
Hardcover ISBN:
9780444522344

About the Author

Andras Guttman

Andras Guttman

András Guttman is an adjunct professor at the Barnett Institute in Northeastern University (Boston, MA), also heading the Horváth Laboratory of Bioseparation Sciences (HLBS) in University of Debrecen (Hungary) and recently received the Marie Curie Chair Professorship from the European Commission. As a separation scientist, his fields of research interest is glycomics, biomarker discovery, and microfabricated device technology. Professor Guttman previously held industrial research positions at Novartis (La Jolla, CA), Genetic Biosystems (San Diego, CA), and Beckman Instruments (Fullerton, CA). He has contributed more than 200 scientific publications, 30 book chapters, edited several textbooks, and holds 18 patents. He is the president of the Hungarian Chapter of the American Chemical Society, board member of CASSS, and on the editorial panels of numerous leading international scientific journals such as Analytical Chemistry, Journal of Chromatography, and Electrophoresis. Dr. Guttman graduated from the University of Veszprem (Hungary) in chemical engineering, where he also received his Ph.D. He has received numerous recognitions such as the Analytical Chemistry Award of the Hungarian Chemical Society in 2000, and became a member of the Hungarian Academy of Sciences in 2004.

Affiliations and Expertise

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

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