Modern Analytical Ultracentrifugation

There are numerous examples in the history of science when the parallel develop ments of two or more disciplines, methodologies, technologies or theoretical in sights have converged to produce significant scientific advances. The decades following the 1950s have produced several such significant advances, as a result of a convergence of developments in molecular biology and in solid state-based electronics instrumentation. Since one of these areas of significant advancement, analytical ultracentrifu gation, has been undergoing a renaissance, we thought it would be a useful activity to call upon a group of researchers who have been developing either the experi mental or theoretical aspects of the methodology and gather in one place a group of articles summarizing the current status of the field. The success of recombinant DNA methodologies at producing biologically active macromolecules of commer cial interest has evoked interests in mechanisms of function. Pursuit of the related questions has emphasized the importance of studies of macromolecular binding and interaction. Several contributions to this volume remind us that analytical ultra centrifugation is rigorously based on solid thermodynamic theory and, as such, is fully capable of providing comprehensive quantitative descriptions of molecular interactions in solution. Furthermore, a number of the chapters provide examples, along with innovative methods for carrying out these characterizations. The past decade has seen several developments that reflect the rebirth of interest in analytical ultracentrifugation.

Analytical ultracentrifugation is one of the most powerful solution techniques for the study of macromolecular interactions, to define the number and stoichiometry of complexes formed, and to measure affinities ranging from very strong to very weak and repulsive. Building on the data analysis tools described in the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles, and the experimental and instrumental aspects in the first volume Basic Principles of Analytical Ultracentrifugation, the present volume Sedimentation Velocity Analytical Ultracentrifugation: Interacting Systems is devoted to the theory and practical data analysis of dynamically coupled sedimentation processes. This volume is designed to fill a gap in biophysical methodology to provide a framework that builds on the fundamentals of the highly developed traditional methods of analytical ultracentrifugation, updated with current methodology and from a viewpoint of modern applications. It will be an invaluable resource for researchers and graduate students interested in the application of analytical ultracentrifugation in the study of interacting systems, such as biological macromolecules, multi-protein complexes, polymers, or nanoparticles.

Analytical ultracentrifugation has become an increasingly important technique for monitoring the size and shape of biological macromolecules. Analytical Ultracentrifugation: Techniques and Methods contains contributions from experts in the field, bringing together the multitude of developments that have taken place in instrumentation and analysis over the past decade into a single volume. This book covers the latest methods in analysis along with an extensive introduction for the novice user. Analysis methods in both sedimentation velocity and sedimentation equilibrium are discussed at length. Protein, protein/DNA, membrane proteins and polymer systems are also explored, along with software developments and non-ideality.

The basis for this volume is the 11th Symposium on Analytical Ultracentrifugation held in March 25-26, 1999 at the University of Potsdam, Germany. This book presents a comprehensive collection of 33 contributions from leading scientists in this field including: Technical and methodological innovations.- Innovations in data analysis.- Hydrodynamics/Modelling.- Synthetic polymers, colloids and supramolecular systems.- Biological systems.- Interacting systems and assemblies. In contrast to the increasing significance of analytical ultracentrifugation, related modern books are very rare. Therefore, this volume will be a helpful source of information to anyone who wants to catch up with the most recent developments and results related to this important analytical method.

This is the first book of its kind to appear for nearly two decades and gives as comprehensive a coverage as is possible of the present state-of-the-art.

This book introduces analytical ultracentrifugation (AUC) as a whole, covering essential theoretical and practical aspects as well as its applications in both biological and non-biological systems. Comprehensive characterizations of macromolecules in a solution are now routinely required not only for understanding the solution system but also for producing a solution with better properties. Analytical ultracentrifugation is one of most powerful and reliable techniques for studying the biophysical behavior of solutes in solution. In the last few years, there have been steady advances made in hardware, software, and applications for AUC. This book provides chapters that cover everything essential for beginners to the most advanced users and also offer updated knowledge of the field on advances in hardware, software, and applications. Recent development of hardware described in this book covers new detection systems that give added dimensions to AUC. Examples of data analysis with essential theoretical explanations for advanced and recently updated software are also introduced. Besides AUC of biological systems including membrane proteins and biopharmaceuticals, AUC applications for non-biological questions are included. AUC studies under non-ideal conditions such as highly concentrated solutions and solutions with high salt concentration are also included. The contributors to this book are leading researchers in the fields of solution biophysics and physical chemistry who extensively employ AUC analysis for their research. From this published work, one can gain new and comprehensive knowledge of recent AUC analysis.

Analytical Ultracentrifugation, the latest volume in Methods in Enzymology, focuses on analytical ultracentrifugation. The scope of this technique has greatly expanded in recent years due to advances in instrumentation, algorithms and software. This volume describes the latest innovations in the field and in the applications of analytical ultracentrifugation in the analysis of macromolecules, macromolecular assemblies, and biopharmaceuticals. Timely contribution that describes a rapidly changing field Leading researchers in the field Broad coverage: instrumentation, basic theory, data analysis, and applications

This book is divided into chapters covering instrumentation, sedimentation velocity runs, density gradient runs, application examples and future developments. In particular, the detailed application chapter demonstrates the versatility and power of AUC by means of many interesting and important industrial examples. Thus the book concentrates on practical aspects rather than details of centrifugation theory.