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 Mechanisms Underlying Protein Sorption and Transport Within Polysaccharide based Stationary Phases for Ion exchange Chromatography written by James Michael Angelo and published by . This book was released on 2016 with total page 229 pages. Available in PDF, EPUB and Kindle. Book excerpt: Ion-exchange chromatography is a powerful tool that is widely utilized in the downstream purification of biomolecules. In the biotechnology industry, resins that exhibit enhanced chromatographic performance are widely desired to streamline purification processes and meet production requirements. Polysaccharide-based ion-exchange adsorbents meet many performance standards by possessing high binding capacities and enhanced uptake rates of protein, while being highly compatible with biologics and remaining relatively cost-efficient. The characterization of these materials is crucial for informed decisions during stationary-phase selection and process design. The structural characteristics of agarose-based, dextran-modified agarose-based and cellulosic ion-exchange materials were determined using methods to gauge the pore dimensions and the effect of ionic strength on intraparticle architecture. Inverse size exclusion chromatography (ISEC) analysis and electron microscopy techniques were used to quantitatively assess and visualize the pore structure, respectively. Adsorption behavior in these materials was characterized using methods to evaluate the dynamics of protein uptake as a function of ionic strength and protein concentration using several model proteins. Fast uptake rates were observed in both batch kinetics experiments and time-series confocal laser scanning microscopy (CLSM) for the cellulosic media, suggesting low intraparticle transport resistances relative to external film resistance, even at higher bulk protein concentrations where the opposite is typically observed. While polysaccharide-based materials possess easily accessible microstructures, elution has sometimes been observed to be undesirably slow. In order to determine which physicochemical phenomena control elution behavior, the materials were characterized by their uptake and elution profiles at various conditions. In general, effective elution rates decreased with the reduction of accessible pore volume. The use of protein stabilizing agents was explored in systems presenting atypical elution behavior. Incorporation of excipients into eluent buffer gave rise to faster elution and significantly lower pool volumes in elution from polymer-modified agarose adsorbents. Finally, thermodynamic modeling based on molecular theory was used in conjunction with single-molecule total internal reflection fluorescence microscopy (TIRFM) to further develop the understanding of fundamental mechanisms of protein adsorption within polymer-modified surface layers. These molecular methods revealed trends and characteristics that were in agreement with protein transport in stationary phase analogues to these surfaces.

Download or read book Ion Exchange Chromatography and Related Techniques written by Pavel Nesterenko and published by Elsevier. This book was released on 2023-11-15 with total page 646 pages. Available in PDF, EPUB and Kindle. Book excerpt: Ion-Exchange Chromatography and Related Techniques defines the current state-of-the-art in ion-exchange chromatography and related techniques and their implementation in laboratory and industrial practice. This book provides a compact source of information to facilitate the transfer of knowledge and experience acquired by separation science specialists to colleagues from diverse backgrounds who need to acquire fundamental and practical information to facilitate progress in research and management functions reliant on information acquired by separation. Individual chapters written by recognized experts lending credibility to the work will allow this book to serve as a high value reference source of current information for analytical and biopharmaceutical chemists. Includes individual chapters written by recognized authoritative and visionary experts in the field to provide an overview and focused treatment of a single topic Presents comprehensive coverage of ion-exchange techniques from theory, to methods, to selected applications for ions and biopolymers Provides Tables and diagrams with commonly used data to facilitate practical work, comparison of results and decision-making

Download or read book Protein Adsorption and Transport in Agarose and Dextran grafted Ion exchange Media written by Antonio R. Ubiera and published by . This book was released on 2005 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book New Bioprocessing Strategies Development and Manufacturing of Recombinant Antibodies and Proteins written by Bob Kiss and published by Springer. This book was released on 2018-12-06 with total page 473 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book review series presents current trends in modern biotechnology. The aim is to cover all aspects of this interdisciplinary technology where knowledge, methods and expertise are required from chemistry, biochemistry, microbiology, genetics, chemical engineering and computer science. Volumes are organized topically and provide a comprehensive discussion of developments in the respective field over the past 3-5 years. The series also discusses new discoveries and applications. Special volumes are dedicated to selected topics which focus on new biotechnological products and new processes for their synthesis and purification. In general, special volumes are edited by well-known guest editors. The series editor and publisher will however always be pleased to receive suggestions and supplementary information. Manuscripts are accepted in English.

Download or read book Preparative Chromatography for Separation of Proteins written by Arne Staby and published by John Wiley & Sons. This book was released on 2017-03-06 with total page 577 pages. Available in PDF, EPUB and Kindle. Book excerpt: Preparative Chromatography for Separation of Proteins addresses a wide range of modeling, techniques, strategies, and case studies of industrial separation of proteins and peptides. • Covers broad aspects of preparative chromatography with a unique combination of academic and industrial perspectives • Presents Combines modeling with compliantce useing of Quality-by-Design (QbD) approaches including modeling • Features a variety of chromatographic case studies not readily accessible to the general public • Represents an essential reference resource for academic, industrial, and pharmaceutical researchers

Download or read book Protein Purification written by Jan-Christer Janson and published by John Wiley & Sons. This book was released on 2011-03-22 with total page 556 pages. Available in PDF, EPUB and Kindle. Book excerpt: The authoritative guide on protein purification—now completely updated and revised Since the Second Edition of Protein Purification was published in 1998, the sequencing of the human genome and other developments in bioscience have dramatically changed the landscape of protein research. This new edition addresses these developments, featuring a wealth of new topics and several chapters rewritten from scratch. Leading experts in the field cover all major biochemical separation methods for proteins in use today, providing professionals in biochemistry, organic chemistry, and analytical chemistry with quick access to the latest techniques. Entirely new or thoroughly revised content includes: High-resolution reversed-phase liquid chromatography Electrophoresis in gels Conventional isoelectric focusing in gel slabs and capillaries and immobilized pH gradients Affinity ligands from chemical and biological combinatorial libraries Membrane separations Refolding of inclusion body proteins from E. coli Purification of PEGylated proteins High throughput screening techniques in protein purification The history of protein chromatography

Download or read book Effects of Ionic Strength and Stationary Phase on Protein Adsorption and Transport in Ion exchange Particles written by Stephen R. Dziennik and published by . This book was released on 2002 with total page 612 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Thermodynamic studies of the purification of polymer modified proteins with ion exchange chromatography written by Tim Blaschke and published by . This book was released on 2012 with total page 159 pages. Available in PDF, EPUB and Kindle. Book excerpt:

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 The Kinetics of Protein Adsorption on Ion Exchange Resin written by Lian-Huey Liu and published by . This book was released on 1991 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Ion Exchange Chromatography of Proteins written by Shuichi Yamamoto and published by CRC Press. This book was released on 1988-04-14 with total page 438 pages. Available in PDF, EPUB and Kindle. Book excerpt: Showing chemists how to predict ion-exchange chromatography (IEC) separation behavior and how to determine appropriate operating conditions, this reference illustrates procedures, apparatus, and types of ion exchangers, emphasizing the design and application of large-scale IEC. Complete with more than 100 useful tables and diagrams, Ion-Exchange Chromatography of Proteins explains the effects of each variable on separation behavior ... compares models and equations describing separation behavior of proteins in IEC with experimental results ... introduces design calculation procedures for scale up, adapting IEC for individual requirements ... discusses parameters affecting separation behavior, including adsorption equilibria, stationary phase diffusion, and axial dispersion ... provides examples of the most up-to-date applications, such as high- or medium-performance IEC and large-scale operations ... and reviews both theoretical and experimental literature. Ion-Exchange Chromatography of Proteins serves as an important reference for analytical, agricultural, food, and pharmaceutical chemists, chromatographers, food scientists and technologists, biochemists, and biotechnologists. Book jacket.

Download or read book Dissertation Abstracts International written by and published by . This book was released on 2008 with total page 1006 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Chromatography written by E. Heftmann and published by Elsevier. This book was released on 2004-04-16 with total page 706 pages. Available in PDF, EPUB and Kindle. Book excerpt: Chromatography has emerged as the most important and versatile analytical method. The book is not only an updated version of Heftmann's classical text, but it covers areas of future importance, such as microfluidics and computer resources. Under his experienced guidance, authorities in each field have contributed their practical experience to an integrated treatment of modern micro analysis. Part B of this two volume set brings the traditional field of application up to date. These include amino acids and proteins, nucleic acids and their constituents, lipid, and carbohydrates. Special chapters are devoted to the most important areas of application: drug and environmental analysis. Forensic and phytochemical applications are covered for the first time. Together with an overview of computer resources, the subject index allows novices as well as experts to obtain rapid and authoritative guidance to analytical problems, such as choice of methods and optimization of techniques and instrumentation. 1. Each chapter written by an authority 2. Thorough treatment of the theoretical basis of separation methods 3. Practical guide for performing analyses

Download or read book Polymers as Biomaterials written by W. Shalaby and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 384 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nearly 4000 years ago, the Egyptians used linen, a natural polymeric material, for suturing wounds. About 600 B.C., the Indians used other forms of natural polymers such as cotton, horse hair, and leather in repairing wounds. Wound closure procedures using silk sutures, based mostly on polypeptides, are likely to have been practiced during the second century. Surgical application of natural polymers continued to represent the major use of polymers until the twentieth century. Not too long after the development of several major synthetic polymers, their use in biomedical applications has attracted the attention of many re searchers and clinicians. Over the past few years, interest in the biomedical applications of polymers has grown considerably. This has been the result of the inevitable collaborative efforts of in novative materials scientists, engineers and clinicians. The es tablishment of the Society for Biomaterials, in our opinion, cata lyzed the growing interest in the use of polymers for biomedical application. In a major effort to bring team players even closer, a five-day symposium on "Polymers as Biomaterials" was held in Seattle, Washing ton, in March, 1983 as part of the national meeting of the American Chemical Society. The symposium was designed to provide a forum for communicating technical and clinical data to colleagues with a broad spectrum of interest in the biomedical applications of polymers.

Download or read book Structure and Dynamics of Confined Polymers written by John J. Kasianowicz and published by Springer Science & Business Media. This book was released on 2002-07-31 with total page 46 pages. Available in PDF, EPUB and Kindle. Book excerpt: Polymers are essential to biology because they can have enough stable degrees of freedom to store the molecular code of heredity and to express the sequences needed to manufacture new molecules. Through these they perform or control virtually every function in life. Although some biopolymers are created and spend their entire career in the relatively large free space inside cells or organelles, many biopolymers must migrate through a narrow passageway to get to their targeted destination. This suggests the questions: How does confining a polymer affect its behavior and function? What does that tell us about the interactions between the monomers that comprise the polymer and the molecules that confine it? Can we design and build devices that mimic the functions of these nanoscale systems? The NATO Advanced Research Workshop brought together for four days in Bikal, Hungary over forty experts in experimental and theoretical biophysics, molecular biology, biophysical chemistry, and biochemistry interested in these questions. Their papers collected in this book provide insight on biological processes involving confinement and form a basis for new biotechnological applications using polymers. In his paper Edmund DiMarzio asks: What is so special about polymers? Why are polymers so prevalent in living things? The chemist says the reason is that a protein made of N amino acids can have any of 20 different kinds at each position along the chain, resulting in 20 N different polymers, and that the complexity of life lies in this variety.

Download or read book Simulated Moving Bed Technology written by Alirio Rodrigues and published by Butterworth-Heinemann. This book was released on 2015-05-23 with total page 305 pages. Available in PDF, EPUB and Kindle. Book excerpt: Simulated Moving Bed Technology is one of only a few books solely dedicated to SMB. It aims to equip you with the tools and skills needed for SMB modelling, simulation, design and operation. The theory presented is supported by real-world examples from the petrochemical, sugar and pharma industries. The core of the book focuses on SMB process development and model validation. It explains basic process concepts and definitions, fundamental principles and equations, as well as typical applications and recent advances made to address challenges. Its specialist content makes it a must have book for people working with SMB technology, chemical engineering post-graduate students and process and R&D engineers in industry. Will provide you with the tools and predictive approaches for use in SMB design and performance assessment Covers SMB modelling, simulation, design and operation as applied to separation and reaction processes Discusses process intensification through coupling SMB and reaction in SMBR, membrane permeation and SMBR in PermSMBR and introduces the design of SCPC