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A mixture of two polymers, or one polymer and a salt, in an aqueous medium separates into two phases: this phenomenon is useful in biotechn- ogy for product separations. Separation of biological molecules and particles in these aqueous two-phase systems (ATPS) was initiated over 40 years ago by P.-Å. Albertsson, and later proved to be of immense utility in biochemical and cell biological research. A boost in the application of ATPS was seen when problems of separations in biotechnology processes were encountered. Its simplicity, biocompatibility, and amenability to easy scaleup operations make the use of ATPS very attractive for large-scale bioseparations. Despite the advantages ATPS enjoys over other separation techniques, the application of two-phase systems has for a long time been confined to selected labora- ries. Recent years have, however, shown a trend in which increasing numbers of researchers employ two-phase partitioning techniques in both basic and applied research.
This comprehensive and unique text presents a full overview of downstream processing useful for those new to the concept as well as professionals with experience in the area. The history and theoretical principles of Aqueous Two-Phase Systems (ATPS) are covered in depth. Information on ATPS characterization and application is included, and ATPS equilibria and system parameters that have significant effect on partition behavior are studied. Aqueous Two-Phase Systems for Bioprocess Development for the Recovery of Biological Products addresses specific applications of ATPS for the recovery and partial purification of high molecular weight compounds such as proteins, nucleic acids and polysaccharides, particulate bioproducts such as cells and organelles and low molecular weight compounds. Non-conventional strategies involving ATPS such as affinity systems, continuous liquid-liquid fractionation stages and the recovery from plant extracts are presented. Economic analysis of the application of ATPS in comparison to other fractionation techniques, particularly liquid chromatography, is considered, as are opportunity and current trends in the ATPS research area. Each chapter utilizes the contributors' experimental expertise in traditional and non-conventional ATPS strategies, as well as analysis of areas of opportunity and perspectives on the development and future applications of ATPS in both the lab and larger scale operations. The result is a thorough and singular overview of ATPS which has not been matched by any other text on the market.
Partitioning in Aqueous Two-Phase Systems: Theory, Methods, Uses, and Applications to Biotechnology is a collection of papers that discusses the applications of aqueous two-phase systems to problems of separation and extraction of macromolecules, organelles, and cells. Papers focus on the theoretical basis and the practical details of the procedures used. Some of the papers describe in one or a few steps how two components can be separated by the investigator manipulating their partitions so that one component is in one phase and the other component is in the other phase or at the interface. Investigators can also avail of developed batch extractions for plant organelles, cell membranes, nucleic acids, and proteins. The book cites as an example the partitioning of right-side-out and inside-out vesicles (obtained from fragments of thylakoid membranes) to the top and bottom phases, respectively, of a Dx-PEG system. Other papers describe the use of the countercurrent distribution when single extraction steps are not sufficient to produce a separation in materials that do not differ greatly in their partitioning behavior. The collection can prove valuable for bio-chemists, cellular biologists, micro-biologists, and developmental biologists.
Aqueous Two-Phase Systems (ATPS) have been used since their discovery for the isolation of a large number of different biological materials, such as animal and plant cells, microorganisms, viruses, cellular organelles such as chloroplasts, mitochondria, membrane vesicles, and also in the purification of proteins and nucleic acids. In Aqueous Two-Phase Systems: Properties, Functions and Advantages, the authors begin by discussing the benefits of these systems.The following chapter presents the main forms of the purification of lipases from aqueous biphasic systems, addressing and discussing the fundamentals for the formation, use and optimization of these systems applied in the biotechnological context. The authors address the basis of aqueous-biphasic systems, aqueous two-phase systems for enzymatic purification, purification of lipases and combined uses of aqueous two-phase systems.The most important experimental parameters that affect the efficiency of metal ions extraction in aqueous PEG-based two-phase systems are examined to provide a starting point in the design of a suitable system for the extraction of metal ions. Compared to the conventional solvent extraction, which uses toxic, flammable and volatile organic solvents and can be quite expensive, the extraction in aqueous PEG-based two-phase systems is considered a more environmental friendly and economically viable method.Also in this compilation, an overview about application of ATPSs in microstructured devices is provided. Microstructured devices offer potential benefits due to well-defined high specific interfacial areas available for heat and mass transfer. These areas increase transfer rate and enhance yield, selectivity and process control. The application of ATPSs in Micro Total Analysis Systems is described as well.
Covers the fundamental principles of solute partitioning in aqueous two-phase systems, explains their important practical features, and furnishes methods of characterization. The information provided by the partition behaviour of a solute in an aqueous two-phase system is examined.
This book offers comprehensive information on the fundamentals and applications of ionic-liquid-based aqueous biphasic systems, which have predominantly (and successfully) been employed as alternative platforms for the extraction, separation and purification of diverse high-value products. The book consists of an initial introduction providing a brief overview, from fundamentals to applications, followed by nine chapters addressing the respective phase diagrams (interpretation and characterization) and remarkable examples of their applications. It also includes two final chapters focusing on recent developments in the search for more environmentally-benign and biocompatible ionic-liquid-based aqueous biphasic systems, and on the progress made to date concerning the recovery, recycling and reuse of the phase-forming components, the goal being the development of cost-effective and sustainable processes. The book offers an interesting and useful guide for a broad readership in the fields of green chemistry, biotechnology, chemical engineering, and biochemistry, among others. Mara G. Freire is a Coordinator Researcher at CICECO - Aveiro Institute of Materials, Chemistry Department, University of Aveiro, Portugal.
"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.
Giant vesicles are widely used as a model membrane system, both for basic biological systems and for their promising applications in the development of smart materials and cell mimetics, as well as in driving new technologies in synthetic biology and for the cosmetics and pharmaceutical industry. The reader is guided to use giant vesicles, from the formation of simple membrane platforms to advanced membrane and cell system models. It also includes fundamentals for understanding lipid or polymer membrane structure, properties and behavior. Every chapter includes ideas for further applications and discussions on the implications of the observed phenomena towards understanding membrane-related processes. The Giant Vesicle Book is meant to be a road companion, a trusted guide for those making their first steps in this field as well as a source of information required by experts. Key Features • A complete summary of the field, covering fundamental concepts, practical methods, core theory, and the most promising applications • A start-up package of theoretical and experimental information for newcomers in the field • Extensive protocols for establishing the required preparations and assays • Tips and instructions for carefully performing and interpreting measurements with giant vesicles or for observing them, including pitfalls • Approaches developed for investigating giant vesicles as well as brief overviews of previous studies implementing the described techniques • Handy tables with data and structures for ready reference
Contributors from European and US universities and corporations review recent developments in the integration of downstream processing and bioconversion, describing their experience with many separation technologies, including some still in the experimental stage. The topics include the construction
Downstream bioprocesses have a significant role to play in the creation of a sustainable bio-based economy, enabling the creation of new products and systems from the more sustainable bioprocessing of natural products. Liquid Biphasic System: Fundamentals, Methods, and Applications in Bioseparation Technology explores in detail the fundamental processes and applications of this new separation system, aiding understanding of the basic principles of the technique and offering constructive criticisms on the latest findings.Including coverage of the background, principles, mechanisms, and applications, Liquid Biphasic System addresses how to adapt the technology for the purification of useful compounds with greater cost efficiency and greener processing. It is essential reading for bioprocess engineers, biochemical engineers, biosystem engineers, chemists and microbiologists working in the fields of bioprocessing. Researchers, scientists, and engineers concerned with the selection and evaluation of alternative bioseparation processes will find the book particularly useful. - Provides information and examples of advanced separations in a single source - Includes detailed descriptions of novel bioseparation systems - Covers the latest technologies related to advanced liquid–liquid separation and their applications in various industries