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Biomolecular and Bioanalytical Techniques
book

Biomolecular and Bioanalytical Techniques

by Vasudevan Ramesh
June 2019
Beginner to intermediate content levelBeginner to intermediate
576 pages
25h 29m
English
Wiley
Content preview from Biomolecular and Bioanalytical Techniques

11 Applications and Complementarity of Analytical Ultracentrifugation and Light‐Scattering Techniques

Chad A. Brautigam

Department of Biophysics, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390‐8816, USA

11.1 Introduction

Analytical ultracentrifugation (AUC) and light scattering (LS) are methods that have a rich history in the characterisation of biological macromolecules [1,2]. For example, AUC was critical to the discovery that proteins exist as discrete species [3,4]. Both techniques were used to probe the solution properties of important, large biomolecular assemblies like haemoglobin [4] and viruses (e.g. [5,6]). In the modern laboratory, advances in instrumentation, methodologies, and analyses have allowed these approaches to be routinely used on biomolecules as small as peptides or even sugars [79].

AUC and LS have many advantages that continue to fuel their popularities. Perhaps foremost among these is that they report on molecular attributes that are critical to understanding biological functioning: molar mass, size, shape, and stoichiometry. However, a practical advantage is that they do not require any kind of labelling or immobilisation; in other words, they are solution techniques that can be deployed on native biomolecules. This is a particular advantage when studying ensembles of macromolecules in the context of their interactions with one another (see Section 11.5.4). Further, AUC and LS are based on easily accessible ...

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