Affinity Chromatography

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.

Bioaffinity chromatography has increasingly become the method of choice for the purification, determination or removal of many biologically active substances.

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 Chromatograph

Matrix preparation and applications; Activation procedures; Cross-linking agents for coupling matrices to spacers; Operational methodologies; Ligands for immobilisation; Quantitative characterisation of interactions by affinity chromatography; Zonal elution quantitative affinity chromatography and analysis of molecular interactions; Cell separation by affinity chromatography ligand immobilisation to solid supports through cleavable mercury-sulphur bonds.

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.

Thirty-eight years after its introduction, affinity chromatography remains a key tool in the armory of separation techniques available to separation and interaction scientists. Expanded and updated from the first edition, Affinity Chromatography: Methods and Protocols, Second Edition, provides the beginner with the practical knowledge needed to develop affinity separations suitable for a variety of applications relevant to the post-genomic era. This second edition expands on the first edition by introducing more state-of-the-art protocols used in affinity chromatography. This new edition also describes protocols that demonstrate the concept of affinity chromatography being applied to meet the modern high throughput screening demands of researchers and development scientists whilst expanding on some more traditional affinity chromatography approaches that have become of greater interest to separation scientists. Chapters in this cutting-edge text expand on affinity chromatography techniques that currently enjoy frequent citation in the literature from those purifying biomolecules. Other chapters include protocols describing the use of a variety of fusion tags as well as how to cleave them, so as to allow the scientists to study the native phenotype of the protein. Renowned researchers also include protocols detailing diverse applications of affinity chromatography such as its use in catalytic reactions, DNA purification, whole cell separations and for the isolation of phosphorylated proteins. Affinity Chromatography: Methods and Protocols, Second Edition, is an essential reference for those interested in separation sciences, particularly in the pharmaceutical and biological research sectors, that have an interest in isolating macromolecules rapidly, quantitatively, and with high purity.

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