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Immobilized Affinity Ligand Techniques

Author: Greg T. Hermanson
Size: 69.69 MB
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This book is a practical guide to the preparation and use of immobilized affinity ligands for purification, catalysis, and analysis. Special emphasis is given to immunochemical techniques including antibody isolation, preparation of antibody fragments using immobilized enzymes, and immunoaffinity chromatography. The book provides easy-to-follow, well-tested protocols to allow the uninitiated to use these techniques to the maximum advantage with minimum hassle. In addition, it shows researchers how to save money by making their own optimized affinity supports. Key Features * Matrix activation * Ligand immobilization * Binding and elution of target molecules * Enzyme catalysis on solid supports * Analytical affinity chromatography * Isolation/purification of antibodies * Preparation of antibody fragments * Immunoaffinity chromatography * Immobilization of nucleic acids * Use of immobilized ligands for removal of trace contaminants Practical advice on choosing: * Matrices * Spacers * Methods of activation and coupling Background information and insights on: * Affinity interactions * The ease and power of affinity chromatography * Attaching molecules to insoluble supports * Matrices currently in use * Over 20 methods of activation * Spacers * Extensive References

Bioconjugate Techniques

Author: Greg T. Hermanson
Publisher: Academic Press
ISBN: 0123822408
Size: 27.58 MB
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Bioconjugate Techniques, 3rd Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions, with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Offers a one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Provides step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates Features full color illustrations Includes a more extensive introduction into the vast field of bioconjugation and one of the most thorough overviews of immobilization chemistry ever presented

Antibody Techniques

Author: Vedpal S. Malik
Publisher: Academic Press
ISBN: 1483285138
Size: 35.56 MB
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The applicability of immunotechniques to a wide variety of research problems in many areas of biology and chemistry has expanded dramatically over the last two decades ever since the introduction of monoclonal antibodies and sophisticated immunosorbent techniques. Exquisitely specific antibody molecules provide means of separation, quantitative and qualitative analysis, and localization useful to anyone doing biological or biochemical research. This practical guide to immunotechniques is especially designed to be easily understood by people with little practical experience using antibodies. It clearly presents detailed, easy-to-follow, step-by-step methods for the widely used techniques that exploit the unique properties of antibodies and will help researchers use antibodies to their maximum advantage. Detailed, easy-to-follow, step-by-step protocols Convenient, easy-to-use format Extensive practical information Essential background information Helpful hints

Handbook Of Affinity Chromatography

Author: Toni Kline
Publisher: CRC Press
ISBN: 1482285010
Size: 52.42 MB
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Outlining the fundamental principles by which all interactions occur, this reference focuses on harnessing the biochemistry of bioorganic compounds in order to separate them, presenting new techniques and applications that affect the planning of research strategies. The contributors discuss how to c

Protein Analysis And Purification

Author: Ian M. Rosenberg
Publisher: Springer Science & Business Media
ISBN: 1461220564
Size: 20.37 MB
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How one goes about analyzing proteins is a constantly evolving field that is no longer solely the domain of the protein biochemist. Investi gators from diverse disciplines find themselves with the unanticipated task of identifying and analyzing a protein and studying its physical properties and biochemical interactions. In most cases, the ultimate goal remains understanding the role(s) that the target protein is playing in cellular physiology. It was my intention that this manual would make the initial steps in the discovery process less time consuming and less intimidating. This book is not meant to be read from cover to cover. The expanded Table of Contents and the index should help locate what you are seeking. My aim was to provide practically oriented information that will assist the experimentalist in benchtop problem solving. The appendices are filled with diverse information gleaned from catalogs, handbooks, and manuals that are presented in a distilled fashion designed to save trips to the library and calls to technical service representatives. The user is encouraged to expand on the tables and charts to fit individual experimental situations. This second edition pays homage to the computer explosion and the various genome projects that have revolutionized how benchtop scientific research is performed. Bioinformatics and In silica science are here to stay. However, the second edition still includes recipes for preparing buffers and methods for lysing cells.

Handbook Of Affinity Chromatography Second Edition

Author: David S. Hage
Publisher: CRC Press
ISBN: 9780824751982
Size: 59.23 MB
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This essential handbook guides investigators in the theory, applications, and practical use of affinity chromatography in a variety of fields including biotechnology, biochemistry, molecular biology, analytical chemistry, proteomics, pharmaceutical science, environmental analysis, and clinical chemistry. The Handbook of Affinity Chromatography reflects upon important factors to consider in the development of affinity methods, such as the choice of support material, immobilization methods, and application or elution conditions. It reviews common affinity methods and explores the latest preparative, analytical, and biophysical applications, including the use of affinity chromatography with other separation techniques and analytical systems. This basis seamlessly supports the discussion of recent developments in techniques including the use of affinity ligands in capillary electrophoresis, mass spectrometry, microanalytical systems, and optical biosensors. New chapters feature expanded discussions on molecularly imprinted polymers and biomimetic ligands, chromatographic immunoassays, affinity-based immunoassays, affinity-based chiral stationary phases, and affinity ligands in multidimensional systems. Engaging the collaboration of 48 scientists and students from 23 laboratories and organizations to present the latest information on affinity methods, the Handbook of Affinity Chromatography illustrates a wide range of applications and theory for scientists, students, and laboratory workers throughout the fields of chemistry and biology.

Immobilized Biochemicals And Affinity Chromatography

Author: R. Dunlap
Publisher: Springer Science & Business Media
ISBN: 1468469827
Size: 47.96 MB
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This volume contains most of the papers presented at the Sym posium on Affinity Chromatography and Immobilized Biochemicals, which was held on November 7-9, 1973 in Charleston, South Carolina in conjunction with the Southeastern Regional American Chemical Society meeting. The topics of the symposium represent two new biochemical frontiers which have emerged in recent years through the ingenious development and application of solid phase biochemi cal technologies. Affinity chromatography involves the use of selected ligands, covalently bound to a solid support such as cel lulose, glass, Sepharose, or polyacrylamide and exploits the princi ple of biochemical recognition between the ligand in the solid phase and a selected macromolecule to facilitate the rapid and often quantitative purification of enzymes, antibodies, antigens, hormones, other proteins, etc. The area of immobilized biochemicals includes the use of coenzymes, oligo- and po1ynuc1eotides, enzymes, and multistep enzyme systems which are immobilized or entrapped in the solid phase. The goals of the symposium were the revi~w of the status of affinity chromatography and immobilized biochemicals, the presentation of new data and ideas in both areas, and the establish ment of a dialogue between research workers in these two evolving disciplines which are so closely interrelated. The papers published in this volume provide the reader with reviews of several topics inherent in the solid phase biochemistry together with a series of timely manuscripts concerning new techniques and applications both in the use of affinity chromatography and in the investigation of immobilized biochemicals.

Bioaffinity Chromatography

Author: J. Turková
Publisher: Elsevier
ISBN: 9780080858630
Size: 56.45 MB
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Bioaffinity chromatography is now the preferred choice for the purification, determination or removal of many biologically active substances. The book includes information on biologically active substances with their affinants, solid supports and methods of coupling, summarized in tables covering classical, high-performance liquid and large-scale bioaffinity chromatography. Optimization of the preparation and the use of highly active and stable biospecific adsorbents is discussed in several chapters. Following a chapter dealing with the choice of affinity ligands, affinity-sorbent bonding is described in detail. Other chapters give information on solid supports, the most common coupling procedures and a general discussion of sorption and elution. Several applications of bioaffinity chromatography are described, e.g. quantitative evaluation of biospecific complexes and many applications in medicine and in the biotechnology industry.


Author: M. A. Vijayalakshmi
Publisher: CRC Press
ISBN: 0203302826
Size: 20.44 MB
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The field of bioseparation, and biochromatography in particular, is advancing very rapidly as our knowledge of the properties of molecules and atomic forces increases. This volume covers the basic principles of biochromatography in detail. It assesses different techniques and includes a large number of applications, providing the reader with a multidisciplinary perspective that gives the insight to master the many chromatographic methods. Biochromatography: Theory and Practice is a valuable tool for graduate and research scientists, technicians, engineers and teachers in a range of fields including biochemistry, biotechnology, biorecognition and chromatography.

Molecular Interactions In Bioseparations

Author: That T. Ngo
Publisher: Springer Science & Business Media
ISBN: 1489918728
Size: 28.74 MB
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Rapid advances in molecular biology have accelerated the production of a great number of protein-based therapeutic agents. The major cost in producing these proteins appears to be associated with their purification from the complex mixture of the crude extract. A major challenge to the protein biochemist and the biochemical engineer is the development of rapid, efficient, and cost-effective purification systems. This volume presents state-of-the-art reviews of current methods used in the purifica tion of biological macromolecules that are based on molecular interactions. Thus, the major emphasis is placed on affinity-related techniques. Part I provides a general introduction to affinity chromatography and includes a chapter describing an interesting new technique called "slalom chromatography" for DNA fractionations. Affinity chromatography using molecules of biological origin as the affinity ligand is covered in Part II. Part III describes the use of a special class of biomolecules, antibodies, as affinity ligands. Affinity chromatography with biomimetic ligands is discussed in Part IV. Newer concepts and their applications in bioseparation are presented in Part V. Part VI covers affinity-related techniques such as affinity-based extracorporeal shunts, affinity electrophoresis, affinity precipitation, and affinity extraction. I would like to express my sincere thanks to all the authors, who are recognized experts in their respective fields, for their cooperation and contributions. I thank the editorial staff of Plenum Press for their professionalism, and Mary Phillips Born, Senior Editor, for her encouragement. The support of my family (Ping and Peilin) made it possible to complete editing this book. That T.