Download or read book Protein Adsorption and Diffusion in Charged Agarose Gels written by Emily Belcher Schirmer and published by . This book was released on 2007 with total page 314 pages. Available in PDF, EPUB and Kindle. Book excerpt: The dissertation describes a new method for measuring protein ion exchange kinetics and new data for the adsorption of three different proteins in anionic agarose gels. The gels were prepared as spherical particles to determine chemical and structural properties and the macroscopic protein adsorption behavior, and as thin slabs supported in a microfluidics chip to observe transient concentration profiles with a light microscope. The combination of these approaches provides deeper insight into the nature of protein transport mechanisms than previously possible. The adsorption of cytochrome c, myoglobin, and hemoglobin was studied macroscopically and microscopically. While all proteins exhibited similarly favorable adsorption isotherms, adsorption kinetics and concentration profiles were quantitatively and qualitatively different. Cytochrome c was adsorbed very quickly and resulted in diffuse concentration profiles, while myoglobin adsorbed much more slowly and resulted in sharp fronts. Hemoglobin exhibited lower rates with diffuse profiles. The effects of ionic strength, pH, and agarose content were studied. A transition of transport mechanism was observed with change in ionic strength and pH indicating that specific binding interactions between the protein and matrix affect protein transport of the protein. With respect to agarose content, transport rates decreased as the agarose content increased as expected due to increase in steric hindrance. Various classical rate models were used to describe the kinetics of protein adsorption. However, only a model that accounts for partitioning of the protein into the gel matrix due to overlapping electric double layers was found to be consistent with the data over broad ranges of conditions.

Download or read book Protein Adsorption and Diffusion in Charged Agarose Gels written by Emily Beth Belcher and published by . This book was released on 2005 with total page 160 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Protein Adsorption and Diffusion in Charged Polyacrylamide Hydrogels written by Shawn Michael Russell and published by . This book was released on 2002 with total page 120 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Environmental Colloids and Particles written by Kevin J. Wilkinson and published by John Wiley & Sons. This book was released on 2007-01-30 with total page 702 pages. Available in PDF, EPUB and Kindle. Book excerpt: This text presents the current knowledge of environmental colloids and includes reviews of the current understanding of structure, role and behaviour of environmental colloids and particles, whilst focussing directly on aquatic systems and soils. In addition, there is substantial critical assessment of the techniques employed for the sampling, size fractionation and characterisation of colloids and particles. Chemical, physical and biological processes and interactions involving colloids are described, and particular attention is paid to quantitative approaches that take account of particle heterogeneity and polydispersity. Presents critical reviews of the state-of-the-art knowledge of environmental colloids Critical assessment of techniques employed for the sampling, size fractionation and characterisation of colloids and particles are given Theoretical and experimental aspects of the methods as well as the required developments and possible recommendations are discussed Each chapter gives a brief introduction general enough for the non-specialist Written by a internationally recognized group of contributors

Download or read book Protein Diffusion in Charged Polyacrylamide Gels and Chromatography Media written by Rebecca Karst Lewus and published by . This book was released on 2000 with total page 342 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Protein Adsorption on Pyridine derivatized Agarose Gels written by Sven Oscarsson and published by . This book was released on 1993 with total page 30 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Methods of Protein Separation written by Nicholas Catsimpoolas and published by Springer Science & Business Media. This book was released on 2013-06-29 with total page 339 pages. Available in PDF, EPUB and Kindle. Book excerpt: This open-end treatise on methods concerning pro tein separation had its beginning in an American Chemical Society symposium entitled "Con temporary Protein Separation Methods" which was held in Atlantic City, New Jersey in September 1974. The purpose of the symposium-and subse quently of the present work-was to review the available modern techniques and underlying principles för achieving one of the very important tasks of experimental biology, namely the separation and characterization of proteins present in complex biological mixtures. Physicochemical characterization was covered only as related to the parent method of fractionation and there fore involved mostly mass transport processes. Additionally, the presentation of methods for gaining insight into complex interacting protein profiles was considered of paramount importance in the interpretation of separation patterns. Finally, specific categories of proteins (e. g. , chemically modified, deriving from a specific tissue, conjugated to different moieties, etc. ) require meticulous trial and selection and/or modification of existing methodology to carry out the desired separation. In such cases, the gained experience provides valuable guidelines for further experimentation. Although powerful techniques exist today for the separation and related physicochemical characterization of pro teins, many biological fractionation problems require further innovations. It is hoped that the description in the present treatise of some of the available separation tools and their limitations will provide the necessary integrated background for new developments in this area. Nicholas Catsimpoolas Cambridge, Massachusetts vü CONTENTS Contents of Volume 1 . xvii Chapter 1 Scanning Gel Cbromatography Gary K. Ackers I.

Download or read book Protein Diffusion in Charged Polyacrylamide Gels written by Rebecca K. Lewus and published by . This book was released on 1999 with total page 2 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Journal of Chromatography written by and published by . This book was released on 2002 with total page 676 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Protein Transport and Adsorption in Polymer modified Ion exchange Media written by Brian D. Bowes and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Ion-exchange chromatography can be a powerful tool for protein purification. New composite ion-exchange resins that combine rigid base matrices with softer polymer layers that have high adsorption capacity and enhanced uptake potential are emerging as attractive options for high-productivity purification processes. However, as the mechanistic origins of the improvements observed for these polymer-modified resins are not fully understood, examination of the effects of the polymer modifiers is warranted. Thus, significant efforts were undertaken to characterize the similarities and differences between a variety of resins with and without polymer modifiers. The characterization included measurements of equilibrium protein adsorption capacities, protein uptake rates, and column breakthrough behavior. In this work, the primary resins used were the traditional agarose-based resin SP Sepharose FF and the dextran-modified agarose resins SP Sepharose XL and Capto S. SP Sepharose FF and SP Sepharose XL made a particularly useful pair of resins to compare because the underlying agarose base matrix is the same for both. Thus, it was anticipated that it would be possible to examine some of the direct consequences of the dextran modification. The role of protein size was also an important consideration here. The small protein lysozyme, the moderate protein lactoferrin, and a large monoclonal antibody were used to test size effects. Experimental techniques were also chosen to study behavior at varied length scales, ranging from the column scale down to the individual-particle scale. Protein adsorption isotherms were measured on a traditional agarose-based resin and on two dextran-modified agarose-based resins. Higher equilibrium capacities were achievable for the dextran-modified resin and seemed to be attributable to volumetric partitioning of protein into the dextran layer compared to the monolayer adsorption on the agarose surface for the traditional resin. For a small protein, isocratic retention experiments revealed that the retention mechanism was quite similar between the two resin classes, suggesting that the dextran does not significantly change the nature of the protein-resin interaction. However, larger proteins were increasingly excluded from the dextran layer as salt concentrations were increased, leading to suppressed equilibrium adsorption capacity at intermediate salt levels. Knowledge of the protein and resin properties were used to rationalize the maximum equilibrium capacities that might be expected on the dextran-modified resins and revealed that the capacities are typically limited by the available volume for adsorption, but that the capacity might be limited by the available resin charge when the protein charge density is high relative to that on the resin. Column breakthrough experiments revealed high achievable dynamic binding capacities for the dextran-modified resins. For both resin classes, dynamic capacities were typically controlled by intraparticle transport limitations at low salt levels, especially for the larger proteins, and were generally limited by equilibrium capacities at higher salt levels. However, the transition between the intraparticle uptake control and the equilibrium capacity control occurred at different salt levels between the resin classes, suggesting a fundamental difference in the balance of adsorption and transport in the resins. Likewise, confocal microscopy experiments showed that, in spite of the similar protein retention properties, uptake was significantly enhanced on the dextran-modified resins, again reflecting a difference in the interplay between protein adsorption and transport. Equilibrium capacity and intraparticle diffusivity information were used to compare predictions from limiting models with experimental breakthrough data. While the agreement was good for the traditional resin, significant discrepancies were observed for the dextran-modified resins. The primary feature hypothesized to be lacking from the limiting models was a slow approach to equilibrium caused by an initially inefficient packing of protein within the dextran layer that gradually improves over time. A conceptual model accounting for this behavior was developed and showed some success in capturing the correct experimental behavior. Additionally, custom variants were used to examine the effects of the resin dextran content and charge density on protein transport and adsorption. Isocratic retention experiments revealed that increasing the dextran content could increase the severity of the high-salt exclusion of protein from the dextran, but that increasing the charge density provides an increased electrostatic driving force that can help overcome this steric exclusion. For the most part, the effect of dextran content was difficult to isolate, but the charge density was shown to correlate well with trends in adsorption capacity, isocratic retention times, and uptake and desorption rates. It was speculated from a number of observations that resins with an intermediate charge density should give rise to the highest achievable dynamic binding capacities and experimental investigations seemed to confirm this hypothesis. Finally, observations were made on a traditional methacrylate-based resin and on a resin with the same base matrix, but modified with proprietary polymers. Some effects of polymer modification similar to those for the agarose-based resins were observed, but some differences were attributable to differences in the resin structures. For example, the greater enhancement in equilibrium capacity with polymer-modification of the methacrylate-based resin could be attributed to the smaller specific surface area of the underlying methacrylate base matrix compared to that of the agarose materials. Additionally, adsorption and transport information seemed to indicate that the polymer-modifiers in the methacrylate-based resin are more subject to swelling and shrinking with changes in salt concentration than are the dextran-modifiers in the agarose media. Thus, while knowledge of the identity of the proprietary polymer could certainly enhance understanding of resin performance, some functional properties and consequences of the polymer were still evident.

Download or read book Bioanalytics written by Friedrich Lottspeich and published by John Wiley & Sons. This book was released on 2018-05-29 with total page 1151 pages. Available in PDF, EPUB and Kindle. Book excerpt: Analytical methods are the essential enabling tools of the modern biosciences. This book presents a comprehensive introduction into these analytical methods, including their physical and chemical backgrounds, as well as a discussion of the strengths and weakness of each method. It covers all major techniques for the determination and experimental analysis of biological macromolecules, including proteins, carbohydrates, lipids and nucleic acids. The presentation includes frequent cross-references in order to highlight the many connections between different techniques. The book provides a bird's eye view of the entire subject and enables the reader to select the most appropriate method for any given bioanalytical challenge. This makes the book a handy resource for students and researchers in setting up and evaluating experimental research. The depth of the analysis and the comprehensive nature of the coverage mean that there is also a great deal of new material, even for experienced experimentalists. The following techniques are covered in detail: - Purification and determination of proteins - Measuring enzymatic activity - Microcalorimetry - Immunoassays, affinity chromatography and other immunological methods - Cross-linking, cleavage, and chemical modification of proteins - Light microscopy, electron microscopy and atomic force microscopy - Chromatographic and electrophoretic techniques - Protein sequence and composition analysis - Mass spectrometry methods - Measuring protein-protein interactions - Biosensors - NMR and EPR of biomolecules - Electron microscopy and X-ray structure analysis - Carbohydrate and lipid analysis - Analysis of posttranslational modifications - Isolation and determination of nucleic acids - DNA hybridization techniques - Polymerase chain reaction techniques - Protein sequence and composition analysis - DNA sequence and epigenetic modification analysis - Analysis of protein-nucleic acid interactions - Analysis of sequence data - Proteomics, metabolomics, peptidomics and toponomics - Chemical biology

Download or read book Determination of Diffusion Coefficients and Adsorption Kinetics for Proteins in Commercial Gel Beads written by Margareta von Sivers and published by . This book was released on 1993 with total page 32 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Protein Purification written by Robert K. Scopes and published by Springer Science & Business Media. This book was released on 2013-04-17 with total page 342 pages. Available in PDF, EPUB and Kindle. Book excerpt: New textbooks at alllevels of chemistry appear with great regularity. Some fields like basic biochemistry, organic reaction mechanisms, and chemical thermodynamics are weil represented by many excellent texts, and new or revised editions are published sufficiently often to keep up with progress in research. However, some areas of chemistry, especially many of those taught at the graduate level, suffer from a reallack ofup-to-date textbooks. The most serious needs occur in fields that are rapidly changing. Textbooks in these subjects usually have to be written by scientists actually involved in the research which is advancing the field. lt is not often easy to persuade such individuals to settime aside to help spread the knowledge they have accumu lated. Our goal, in this series, is to pinpoint areas of chemistry where recent progress has outpaced what is covered in any available textbooks, and then seek out and persuade experts in these fields to produce relatively concise but instructive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In some cases the availability of texts in active research areas should help stimulate the creation of new courses. NewYork CHARLES R. CANTOR Preface to the Second Edition The original plan for the first edition of this book was to title it Enzyme Purification: Princip/es and Practice.

Download or read book Protein Chromatography written by Giorgio Carta and published by John Wiley & Sons. This book was released on 2020-06-02 with total page 440 pages. Available in PDF, EPUB and Kindle. Book excerpt: An all-in-one practical guide on how to efficiently use chromatographic separation methods Based on a training course that teaches the theoretical as well as practical aspects of protein bioseparation to bioprocess professionals, this fully updated and revised new edition offers comprehensive coverage of continuous chromatography and provides readers with many relevant examples from the biopharmaceutical industry. Divided into two large parts, Protein Chromatography: Process Development and Scale-Up, Second Edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. The first part introduces chromatographic theory, including process design principles, to enable the reader to rationalize the set-up of a bioseparation process. The second part illustrates by way of case studies and sample protocols how the theory learned in the first part may be applied to real-life problems. Chapters look at: Downstream Processing of Biotechnology Products; Chromatography Media; Laboratory and Process Columns and Equipment; Adsorption Equilibrium; Rate Processes; and Dynamics of Chromatography Columns. The book closes with chapters on: Effects of Dispersion and Rate Processes on Column Performance; Gradient Elution Chromatography; and Chromatographic Column Design and Optimization. -Presents the most pertinent examples from the biopharmaceutical industry, including monoclonal antibodies -Provides an overview of the field along with design tools and examples illustrating the advantages of continuous processing in biopharmaceutical productions -Focuses on process development and large-scale bioseparation tasks, making it an ideal guide for the professional bioengineer in the biotech and pharma industries -Offers field-tested information based on decades of training courses for biotech and chemical engineers in Europe and the U.S. Protein Chromatography: Process Development and Scale-Up, Second Edition will appeal to biotechnologists, analytical chemists, chromatographers, chemical engineers, pharmaceutical industry, biotechnological industry, and biochemists.

Download or read book Preparative Chromatography written by H. Schmidt-Traub and published by John Wiley & Sons. This book was released on 2020-02-27 with total page 714 pages. Available in PDF, EPUB and Kindle. Book excerpt: The third edition of this popular work is revised to include the latest developments in this fast-changing field. Its interdisciplinary approach elegantly combines the chemistry and engineering to explore the fundamentals and optimization processes involved.

Download or read book Protein Electrophoresis in Clinical Diagnosis written by David Keren and published by CRC Press. This book was released on 2003-09-26 with total page 415 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the publication of High-Resolution Electrophorsesis and Immunofixation 2e, there have been ever-increasing advances in the analyses of proteins, by electrophoresis in particular. Protein Electrophoresis in Clinical Diagnosis shows the changes in both techniques and interpretation, presenting a comprehensive review of serum protein techniques, immunofixation techniques, approaches to pattern interpretation, and pattern interpretation in both cerebralspinal fluid and urine. Conditions associated with Monoclunal Gammopathies are considered, as are the appropriate strategies for their detection. David Keren is well-known as the leader in this field, his work on guidelines becoming the benchmark for all those involved in protein detection in serum and urine. Dr Keren's book will be essential in every laboratory, and read by pathologists, chemical chemists, medical technicians and clinicians (particularly hematologists and oncologists).

Download or read book Isoenzymes written by D. W. Moss and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 266 pages. Available in PDF, EPUB and Kindle. Book excerpt: The increased interest in multiple forms of enzymes that began with the application of new methods of fractionation to preparations of enzymes and other proteins some 25 years ago led quickly to an appreciation that the existence of enzymes in multiple forms, or isoenzymes, is a general phenomenon. The results of pioneering studies and those which followed in the early years of isoenzyme research consisted, not surprisingly, mainly of descriptions of the existence and characteristics of hetero geneity in various enzyme systems. Summaries of these results were provided in books such as J . H. Wilkinson's I soenzymes, the first edition of which appeared in 1965. Some clearer ideas of the nature of the phenomena had become apparent by the time that the second edition of Isoenzymes was called for in 1970, and a limited use of the word isoenzymes itself, to describe only certain of the various categories of enzyme multiplicity then recognized, was already being proposed. Nevertheless, a largely enzyme-by-enzyme oTganization of the contents of the book was still appropriate. Considerable advances, both experimental and conceptual, were made in isoenzyme research in the 1970s, and in 1977 Professor Wilkinson suggested to the present author that these should be taken into account in a joint revision of Isoenzymes. Professor Wilkinson's untimely death put. an end to this project and the present book is therefore the work of a single author.