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Author: Mahmoud F. M. Bahnasy Publisher: ISBN: Category : Capillary electrophoresis Languages : en Pages : 221
Book Description
Capillary electrophoresis (CE) is a fast and high efficiency separation technique based on the differential migration of charged species in an electric field. CE is useful for the separation of a wide range of analytes from small ions to large biomolecules. However, CE separations of proteins are challenging due to the adsorption of protein onto the capillary silica surface. Capillary coatings are the most common way to minimize this adsorption. This thesis focuses on the use of two-tailed cationic surfactant based coatings as means of preventing protein adsorption. Factors affecting the stability of two-tailed cationic surfactant coatings have been investigated. The impact of small i.d. capillaries (5-25) μm on enhanced stability of surfactant bilayer cationic coatings and on the efficiency of separation of basic proteins was studied. Using a dioctadecyldimethylammonium bromide (DODAB) coated 5 μm i.d. capillary, exceptional short term stability (210 consecutive runs) and long term stability (300 runs over a 30 day period) were achieved. The average separation efficiency of four basic model proteins was 1.4-2 millions plates/m. DODAB coatings were stable over a pH range of 3-8 as demonstrated by strong anodic magnitude of electroosmotic flow (EOF) and good EOF reproducibility. Surprisingly, at pH ≥ 9, EOF became less anodic and even became suppressed cathodic. The reason is unclear. Chemical degradation of DODAB at high pH was excluded. Increased vesicle size at high pH and/or accelerated desorption may be involved. A surfactant bilayer/diblock copolymer coating was developed to tune the EOF and prevent protein adsorption. The coating consisted of a DODAB bilayer which served as a strong anchor to the capillary wall and polyoxyethylene (POE) stearate to suppress the EOF. The coating has been applied successfully to the capillary zone electrophoretic separation of basic, acidic and histone proteins, and to capillary isoelectric focusing. The ability to tune the EOF enabled both single-step capillary isoelectric focusing (cIEF) and two-step cIEF to be performed. A strongly suppressed EOF coating provided a linear pH gradient and allowed for the separation of two hemoglobin variants HbA and HbS. Factors affecting the stability and EOF of the developed surfactant bilayer/diblock copolymer coating were studied. The magnitude of the anodic EOF can be tuned by varying the hydrophilic block POE chain length. The hydrophobic block of the diblock copolymer accounts for stability of the coating, with a longer (stearate) block giving the best stability. The sequential coating provided a stable and suppressed EOF over a broad range of pH 3.0-11.5. The EOF was suppressed and anodic at low pH. As the pH increases, the EOF was still suppressed but became cathodic. This reversal in EOF of the sequential coating is consistent with the reported applications of the sequential coating, and the behavior of the underlying DODAB bilayer. The sequential coating shows a good stability in buffers containing up to 20% v/v acetonitrile.
Author: Mahmoud F. M. Bahnasy Publisher: ISBN: Category : Capillary electrophoresis Languages : en Pages : 221
Book Description
Capillary electrophoresis (CE) is a fast and high efficiency separation technique based on the differential migration of charged species in an electric field. CE is useful for the separation of a wide range of analytes from small ions to large biomolecules. However, CE separations of proteins are challenging due to the adsorption of protein onto the capillary silica surface. Capillary coatings are the most common way to minimize this adsorption. This thesis focuses on the use of two-tailed cationic surfactant based coatings as means of preventing protein adsorption. Factors affecting the stability of two-tailed cationic surfactant coatings have been investigated. The impact of small i.d. capillaries (5-25) μm on enhanced stability of surfactant bilayer cationic coatings and on the efficiency of separation of basic proteins was studied. Using a dioctadecyldimethylammonium bromide (DODAB) coated 5 μm i.d. capillary, exceptional short term stability (210 consecutive runs) and long term stability (300 runs over a 30 day period) were achieved. The average separation efficiency of four basic model proteins was 1.4-2 millions plates/m. DODAB coatings were stable over a pH range of 3-8 as demonstrated by strong anodic magnitude of electroosmotic flow (EOF) and good EOF reproducibility. Surprisingly, at pH ≥ 9, EOF became less anodic and even became suppressed cathodic. The reason is unclear. Chemical degradation of DODAB at high pH was excluded. Increased vesicle size at high pH and/or accelerated desorption may be involved. A surfactant bilayer/diblock copolymer coating was developed to tune the EOF and prevent protein adsorption. The coating consisted of a DODAB bilayer which served as a strong anchor to the capillary wall and polyoxyethylene (POE) stearate to suppress the EOF. The coating has been applied successfully to the capillary zone electrophoretic separation of basic, acidic and histone proteins, and to capillary isoelectric focusing. The ability to tune the EOF enabled both single-step capillary isoelectric focusing (cIEF) and two-step cIEF to be performed. A strongly suppressed EOF coating provided a linear pH gradient and allowed for the separation of two hemoglobin variants HbA and HbS. Factors affecting the stability and EOF of the developed surfactant bilayer/diblock copolymer coating were studied. The magnitude of the anodic EOF can be tuned by varying the hydrophilic block POE chain length. The hydrophobic block of the diblock copolymer accounts for stability of the coating, with a longer (stearate) block giving the best stability. The sequential coating provided a stable and suppressed EOF over a broad range of pH 3.0-11.5. The EOF was suppressed and anodic at low pH. As the pH increases, the EOF was still suppressed but became cathodic. This reversal in EOF of the sequential coating is consistent with the reported applications of the sequential coating, and the behavior of the underlying DODAB bilayer. The sequential coating shows a good stability in buffers containing up to 20% v/v acetonitrile.
Author: Philippe Schmitt-Kopplin Publisher: Springer Science & Business Media ISBN: 1588295397 Category : Science Languages : en Pages : 808
Book Description
This book presents a selection of current capillary electrophoresis methods used to separate representative types of molecules and particles and in combination with different detection techniques. It includes practical details which are hard to find elsewhere. The volume is intended for beginners in the field and provides an overview of the technique and a starting point for the exploration of the defined literature on different application topics.
Author: Mark A. Strege Publisher: Springer Science & Business Media ISBN: 159259798X Category : Science Languages : en Pages : 340
Book Description
Throughout the more than 20 years that have followed the beginnings of capillary electrophoresis (CE), its application to the analysis of proteins and peptides has continued to be reliable, versatile, and productive. Over time, CE has matured to become a superb complement to HLPC, and in many cases has also evolved as an automated and quantitative replacement for conventional slab gel electrophoresis methods such as SDS-PAGE and isoelectric focusing. Within Capillary Electrophoresis of Proteins and Peptides, we have assembled contributions from researchers who are applying state-of-the-art CE for protein and peptide analysis, including topics that we believe are of great potential both in the present and for the future. In comparison to traditional separation methods, CE represents a miniaturized analysis technique (especially in its microchip-based format) that is highly dependent upon the basic fundamentals of effective sample recovery and high sensitivity detection. With these issues in mind, Chapters 1–4 describe recently developed approaches for both capillary coatings and analyte detection via laser-induced fluorescence. Since the discipline of biotechnology has established itself as a primary platform for the application of CE to the analysis of proteins and peptides, Chapters 5–7 demonstrate a variety of examples of the specific techniques that have been applied for the development of biopharmaceuticals and their commercialization. The methods covered here include also the analysis of oligosaccharides from glycoproteins.
Author: J G Manjunatha Publisher: Royal Society of Chemistry ISBN: 1837671192 Category : Science Languages : en Pages : 207
Book Description
Describing the importance of surfactants in electrochemical investigations related to biologically and environmentally vital chemicals, this book charts the progression of highly responsive electrochemical sensors using surfactants as a modifying agent in the sensor field. It provides contributed chapters from respected researchers on information concerning the activity of target molecules and electron transfer kinetics arising at the surface of the fabricated sensor materials. Surfactant-based electrochemical sensors are of great interest in the quest to find low-cost, fast and highly responsive sensing devices and one aim of this book is to help improve the competence and knowledge in this newly emerging interdisciplinary research area. Attracting an audience of students, academics, industrialists and engineers, it will interest researchers seeking to use non-toxic sensors in their detection challenges.
Author: S.M. Lunte Publisher: Elsevier ISBN: 0080538711 Category : Science Languages : en Pages : 523
Book Description
The book describes the theory and applications of Capillary Electrophoresis (CE) in the field of pharmaceutical and biomedical analysis. It is targeted towards users who are intimately involved in analytical problems, especially those which involve small samples. This book presents the technique of capillary electrophoresis from the point of view of the serious hands-on use in the field of pharmaceutical and biomedical analysis. An overview of general theory is presented to acquaint the novice with the fundamental principles. A more theoretical approach is taken in the presentation of electrokinetic chromatography. The next chapter discusses advances in column technologies, the preceding chapters having provided a foundation as to how separations occur. In the next three chapters, recognized experts in their fields present fundamentals and state-of-the-art techniques in the areas of optical, electrochemical and mass spectrometric detection. The major focus of the remaining chapters is on applications. This includes the analysis of pharmaceuticals, amino acids and peptides, macromolecules, nucleosides, nucleotides and oligonucleotides. The use of CE for analysis of small ions and separation of biological particles is also discussed. The issue of sample preparation for analysis by CE is addressed, especially as it relates to clinical analysis.
Author: James P. Landers Publisher: CRC Press ISBN: 9780849324987 Category : Medical Languages : en Pages : 918
Book Description
Because new information was discovered at an incredible rate since the publication of the successful first edition of this Handbook, this fully updated second edition covers all areas of interest in the field of capillary electrophoresis (CE). A relatively new technology, CE is a principle method for studying the physicochemical properties of proteins, peptides, and other macromolecules. Where applicable, the 30 chapters provide basic underlying theories as well as application-oriented aspects of each technique.Keep up with all the developments in this growing field with the Handbook of Capillary Electrophoresis, Second Edition - a complete guide to the fundamentals of CE and the latest research. The chapters are organized into five units: Modes: Presents a theoretical development of the basic principles governing separation with several modes, including CEC, and discusses their practical aspects. Analyte: Applies CE to the analysis of a specific class of analytes, including organic and inorganic ions, pharmaceuticals, glycoconjugates, peptides, proteins, and DNA fragments. Fundamental Aspects of CE: Technique-oriented information for the practitioner, including the importance of the sample matrix, on-line preconcentration of samples, modes of detection, and specific aspects of CE data analysis. Applications of CE: Includes single cell analysis, CE in DNA sequencing, CE as a clinical diagnostic tool, identifying and quantifying drugs, and for characterizing interacting species. Specialized Aspects of CE: Discusses interfacing CE with mass spectrometry, high-volume throughput continuous CE, microchip CE, control of EOF, and much more. The Handbook of Capillary Electrophoresis, Second Edition, pulls together diverse areas and applications of CE, resulting in an excellent tool for scientists involved in biotechnology and clinical chemistry, as well as the pharmaceutical, bioscience, chemical, and instrument-manufacturing industries. With an applications-oriented focus, the handbook is also a superb manual for workshops, seminars, and graduate courses in separation science.
Author: Charles Henry Publisher: Springer Science & Business Media ISBN: 1597450766 Category : Science Languages : en Pages : 238
Book Description
Leading chemists and engineers concisely explain the principles behind microchip capillary electrophoresis and demonstrate its use in a variety of biochemical applications, ranging from the analysis of DNA, proteins, and peptides to single cell analysis and measuring the impact of surface modification on flow in microfluidic channels. Since surface chemistry must be carefully considered for optimal operation at this scale, the authors also discuss methods of both adsorbed and covalent surface modification for its control. Fabrication methods for producing microchips with glass, poly(dimethylsiloxane), and other polymers are also provided so that even novices can produce simple devices for standard separations. Microchip Capillary Electrophoresis: Methods and Protocols provides a practical starting point for either initiating research in the field of microchip capillary electrophoresis or understanding the full range of what can be done with existing systems.
Author: Andras Guttman Publisher: Newnes ISBN: 0080931359 Category : Science Languages : en Pages : 391
Book 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. 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
Author: Patrick Camilleri Publisher: CRC Press ISBN: 9780849391279 Category : Medical Languages : en Pages : 586
Book Description
The importance of capillary electrophoresis (CE) as an analytical tool has increased dramatically over the last ten years. It has changed from being an exploratory technique, mainly of academic interest, to one that is applied to solve "real" analytical problems. CE is easily adapted to its various modes of operation, often requiring little more than a change of the buffer solution, and is quickly becoming the preferred technique when analyzing minute amounts of available material. Featuring new chapters on CE analysis of inorganic ions and carbohydrates, the new edition of Capillary Electrophoresis not only presents this method as an academic tool, but also provides applications for solving "real-world" analytical problems. This updated Second Edition reflects the increasing use of CE over the last 10 years, how it is being applied, and the basic theoretical aspects of the separation and detection methodology of CE. Capillary Electrophoresis: Theory and Practice will appeal to students and professionals of analytical chemistry, physical chemistry, biochemistry, and biotechnology and includes suitable experiments designed to be attempted by university or college students, or anyone else wishing to familiarize themselves with CE.
Author: Mahmoud F. M. Bahnasy
Author: Mahmoud F. M. Bahnasy
Author: Philippe Schmitt-Kopplin
Author: Mark A. Strege
Author: J G Manjunatha
Author: 
Author: S.M. Lunte
Author: James P. Landers
Author: Charles Henry
Author: Andras Guttman
Author: Patrick Camilleri