Two-Dimensional Separation for Proteomic Analysis
This chapter describes the development of the two-dimensional separation methods using flow field-flow fractionation (F4) and isoelectric focusing for proteomics utility. The methods described here are the rapid, non-gel-based, on-line, two-dimensional se
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S. Kim R. Williams
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Karin D. Caldwell
Editors
Field-Flow Fractionation in Biopolymer Analysis
Editors Prof. S. Kim R. Williams Laboratory for Advanced Separations Technologies Department of Chemistry and Geochemistry Colorado School of Mines Golden, CO 80401 USA [email protected]
Prof. Karin D. Caldwell Department of Physical and Analytical Chemistry Section of Surface Biotechnology Uppsala University 75123, Uppsala Sweden [email protected]
This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying machines or similar means, and storage in data banks. The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. # 2012 Springer-Verlag/Wien SpringerWienNewYork is a part of Springer Science+Business Media springer.at Typesetting: SPi, Pondicherry, India Printed on acid-free and chlorine-free bleached paper SPIN: 12803026 With 93 Figures Library of Congress Control Number: 2011944221 ISBN 978-3-7091-0153-7 e-ISBN 978-3-7091-0154-4 DOI 10.1007/978-3-7091-0154-4 SpringerWienNewYork
Preface
The collection of analytical techniques suitable for separation and characterization of fragile biopolymers contains, among many others, a group of methods collectively referred to as Field-Flow Fractionation (FFF). Common to these methods is that they are liquid phase elution techniques, in which the separation is executed in open channels unobstructed by solid packing materials, and that they offer a wide resolution range particularly well suited for macromolecules and particles. Recently, these techniques have had a strong upswing in use, especially due to the increased availability of convenient–to-handle commercial instrumentation. The FFF techniques differ from each other in terms of the field chosen to accomplish selectivity, e.g. thermal, gravitational, electrical, etc. Today, the hydrodynamic “flow field” is most commonly used, and hence the present collection of articles focuses extensively, although not exclusively, on a number of attractive applications of flow FFF to problem solving in the biomedical field. The growth of a technique brings with it nonuniformity in terminology. For example, asymmetrical flow FFF is commonly designated as AsFlFFF or AF4. This variation is apparent in the published literature and was purposefully maintained in this book. Chapter 1 describes the theory of flow FFF, both in the symmetric and asymmetric channels presently in use. The evolution and fine-tuning of the technique is discussed in conjunction with the effects of channel dimensions and operating conditions on retention and resolution. Chapter 2 discusses the choice of membrane to serve as sample accumulation wall in the flow FFF channel. The discussion leads to a s
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