244 pages | 120 Color Illus.
A follow-up to the experimental and instrumental aspects described in Basic Principles of Analytical Ultracentrifugation, the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles describes the theory and practice of data analysis. Mathematical models for the sedimentation process and the evolution of detected signals are developed in a comprehensive framework, jointly with the description of current and historical strategies for how to extract from noisy experimental data the physical parameters of interest, such as size, mass, and shape, composition, and polydispersity of sedimenting particles.
The methods are extensively illustrated, and supported with practical applications, as well as cross-references where to find the methods in the public domain software SEDFIT and SEDPHAT. The systems covered are discrete or polydisperse mixtures of sedimenting molecules or particles in dilute solution, such as proteins and other biomolecules and their stable complexes, man-made polymers, and nanoparticles, observed in different optical systems. A useful reference for researchers and graduate students of macromolecular disciplines, these methods form the essential foundation for the analysis of dynamic interacting systems, which are covered in the volume Sedimentation Velocity Analytical Ultracentrifugation: Interacting Systems.
Software referenced in the book is available for download at: https://sedfitsedphat.nibib.nih.gov/default.aspx
"The beauty of this book is that it addresses, in just one place, theory, experiments, and computing in application to analytical ultracentrifugation (AUC) performed with biological and soft matter systems. As the author states, it is the ability to compute routine solutions to the Lamm equation of sedimentation that has transformed the capabilities of modern AUC experiments, and the Lamm equation is the cornerstone of this book. The reader is taken through a comprehensive mathematical tour of how to fit sedimentation coefficient distribution functions c(s) in over 200 pages. Following a recap of the basics at the start, we are presented with the main scenarios of how sedimentation is analyzed. This starts with discrete non-interacting particles and the distributions resulting from these, then it moves to consider the distributions that result from non-diffusing and diffusing particles. There are useful discourses into g*(s) analyses and multi-component distributions, and a neat touch is the inclusion of practical considerations in the fits at the end. There is a summary at the end of how the Lamm equation works. Irrespective of what type of system is being worked with, this authoritative text gives insight of how the appropriate analysis tools work. This is a truly excellent resource for any ultracentrifugation laboratory to consult when needed, with well-focused sections, and I will certainly be using it myself!"
—Steve Perkins, University College London, UK
"Covering modern sedimentation velocity methodologies for a number of possible solution systems, this new book is comprehensive, accurate, and precise. Based on a very strong mathematical background and including brief tutorials, one will be able to use this book as a dictionary of computer calculation-based modern sedimentation velocity methods, and refer to it even in 20 or 30 years."
—Susumu Uchiyama, Osaka University, Japan
"What an epic tour de force this book is! Peter Schuck has delivered us a classy and invaluable documentation of the methods underpinning the modern analysis of sedimentation velocity data. The book eloquently explains the methods and background and provides mathematical rigor at the same time. It will be my key encyclopedia and handbook and one in which I will turn to as my first choice for my students as they learn the method!"
—Danny Hatters, Associate Professor, The University of Melbourne, Australia
"This book is a fantastic resource for those using the powerful technique of sedimentation velocity analytical ultracentrifugation to characterize macromolecules or other particles in solution. Dr. Schuck provides a thorough but approachable overview of the equations underlying sedimentation and allows the reader to peer ‘under the hood’ of the widely used analysis program Sedfit. Starting with explanations of fundamental principles, the book also extends into discussions regarding recent developments in the field such as 2D size-and-shape distribution analysis and multispectral resolution of complicated complex stoichiometries. The volume will be popular with scientists in the pharmaceutical industry or academia interested in gaining a deeper understanding of analytical ultracentrifugation and applying their newly found knowledge to their favorite protein or polymer. I, myself, now have several new experiments that I want to try which were inspired by my reading!"
—Andrew Herr, Associate Professor, Cincinnati Children's Hospital Medical Center, USA
"This book provides an excellent and thorough treatment of sedimentation velocity analysis of macromolecules and particles using the analytical ultracentrifuge. The author’s laboratory has contributed, in no small part, to the recent resurgence of the use of analytical ultracentrifugation in the life sciences, through his development and dissemination of methods and algorithms for data analysis. In particular, new analyses have greatly enriched the biophysical information available from sedimentation velocity data. This volume provides an outstanding review of modern sedimentation velocity analysis including recent advances and the potential that they provide for the analysis of molecules in solution. The coverage of foundation, theory, methods, and data analysis is sufficiently broad ranging to be accessible to readers new to the technique, while being an invaluable reference and source of information on the state of the art for experienced practitioners. This book will be highly useful to all researchers interested in the physical properties of soluble molecules, and will be a vital part of the reference library of all molecular biophysics and structural biology research laboratories."
—Michael Griffin, University of Melbourne, Australia
"This is an outstanding contribution to the field of analytical ultracentrifugation by one of the world’s leading experts. From theory to application, this book is a master class in contemporary sedimentation velocity analyses and an absolute must for all researchers with an interest in macromolecular and nanoparticle hydrodynamics."
—Matthew A. Perugini, La Trobe University, Australia
"'The power and beauty of sedimentation velocity analytical ultracentrifugation rests in its simplicity…' That there is truth to this statement is the direct result of the efforts a large number of dedicated individuals who have worked tirelessly to make analytical ultracentrifugation a pleasure for the rest of us, and Peter Schuck has been at the forefront of the wave of those individuals.
Schuck’s work is wonderful; it is what we used to refer to as a ‘big’ book. It should be studied and slowly digested, not read with the thought of having a quick "aha!" revelation….The first two chapters are meant for biophysical experts and those who wish to become such…I especially enjoyed chapter 3’s emphasis on the heterogeneous nature of pure samples and the expected distributions of non-diffusing, non-interacting particles. While chapter 4 continues with the analysis of non-diffusing particles and their analysis by alternative methods, in chapter 5 we are presented with the real world of diffusing particles. For those who have worked and enjoyed their way through the theoretical sections but have limited hands on experience with Analytical Ultracentrifugation, chapter 8 is a welcome must.
While not saying so explicitly, Schuck suggests that it is necessary to have faith in your computer but above all to look at the data - advice that should be ingrained in every experimental scientific endeavor!"
—Jack A. Kornblatt, Assistant Professor Emeritus, Biology, Concordia University, USA
Basic Analysis Principles
Sedimentation of Discrete Non-Interacting Particles
Properties of Sedimentation Coefficient Distributions
Distributions Of Non-Diffusing Particles
Distributions of Diffusing Particles
Sedimentation Coefficient Distributions from Boundary Derivatives and Extrapolations
Practical Analysis of Non-Interacting Systems
Appendix A: Numerical Solutions of the Lamm Equation
Appendix B: Calculating Distributions