Affinity Chromatography And Biological Recognition

Affinity Chromatography and Biological Recognition contains manuscripts presented at the Fifth International Symposium on Affinity Chromatography and Biological Recognition convened in June 12-17, 1983, at St. John's College in Annapolis, Maryland. Organized into six parts encompassing 82 chapters, the book begins by examining the growing synergism between affinity methods and the understanding and study of basic principles of biological recognition. The book then focuses on the trends and progress in the design and application of affinity methods for isolation, therapeutics, diagnostics, and biotechnology. Significant chapters are devoted to the contributions of affinity methodology in such areas as cell membrane receptors, quantitative properties of macromolecular interactions, microscale analytical and preparative applications of high performance affinity chromatography, antibodies as in vivo and in vitro diagnostic and therapeutic agents, and drug targeting. This volume will be a stimulus for broad and creative application of affinity concepts and methods in many fields of biomedical research and biotechnology.

Affinity chromatography, with its exquisite specificity, is based upon molecular recognition. It is a powerful tool for the purification of biomolecules. In recent years, numerous new applications and modified techniques have been derived from gro- specific interactions and biological recognition principles. An up-to-date review of the past, current, and future applications of affinity chromatography has been presented in the introductory chapter by Meir Wilchek and Irwin Chaiken. Though many of these new applications and techniques are well documented in the literature, it is often difficult to find methods that are written with the intent of helping new practitioners of affinity chromatography. This volume on Affinity Chromatography: Methods and Protocols is intended for the novice, as well as for - perts in the field. The protocols are written by experts who have developed and/or successfully employed these methods in their laboratories. Each chapter describes a specific technique, and since the book is intended to help the beginner, each technique is described simply and clearly, making sure that all relevant steps are included, assuming no previous knowledge. Each chapter contains an introduction describing the principles involved, followed by a Materials and Methods section, which lays the groundwork for the reader to conduct experiments step-by-step, in an orderly fashion. The following Notes section, which describes many of the problems that can occur, makes suggestions for overcoming them, and provides alternate procedures. These are precisely the sort of important, practical details that never seem to appear in the published literature.

Most will agree that one major achievement in the bio-separation techniques is affinity chromatography. This coined terminology covers a myriad of separation approaches that relies mainly on reversible adsorption of biomolecules through biospecific interactions on the ligand. Within this book, the authors tried to deliver for you simplified fundamentals of affinity chromatography together with exemplarily applications of this versatile technique. We have always been endeavor to keep the contents of the book crisp and easily comprehensive, hoping that this book will receive an overwhelming interest, deliver benefits and valuable information to the readers.

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 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.

This volume presents discussions of theoretical and experimental considerations that have led to the analytical affinity chromatography field, as well as current efforts to use this methodology to characterize the interaction mechanisms of biological macromolecules and to establish conditions for employing bioaffinity chromatographic systems as preparative tools. The chapters include a comprehensive discussion of interactive chromatography theory (DeLisi and Hethcote), a review of experimental data obtained for biological macromolecules and the relevant theoretical considerations of affinity chromatography which led to them (Swaisgood and Chaiken), an evaluation of rate processes in affinity chromatography and the potential to determine biologically meaningful chemical rate constants (Walters), and the use of quantitative and molecular considerations to design affinity chromatographic systems (Stellwagen and Liu).

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