High Resolution Chromatography: A Practical Approach

Information given in this post is taken from the preface of this book. In my opinion, this is one of the best books out there in the market for the advance chromatographic techniques. At the bottom of the post you will find the download link, I think it might be useful for you for further review and information. Remember, that link is provided for education purpose only.

This is particularly the case with ‘non-specific’ separations based on the physico-chemical aspects of the macromolecules to be resolved, such as charge, size, and hydrophobicity (Chapter 1) and which often represent the initial clean-up step, prior to the application of affinity, or other specific separations. In addition, the development of new and increasingly sophisticated detector technologies (Chapter 3) now allows much more information, concerning the separated macromolecules emerging from a chromatographic column, to be determined. In some cases, specialized chromatographic ‘niches’, such as microbore chromatography, which allow the resolution of extremely small amounts of sample to be performed, require special attention to chromatographic parameters that could normally be taken for granted in laboratory scale preparations. The application of microscale separation procedures is discussed in detail in Chapter 2 by Yang. Finally, the technology of capillary electrophoresis and allied techniques is covered by Hu and Martin (Chapter 4). Although not strictly chromatographic, capillary electrophoretic techniques are often used in conjunction with high resolution chromatographic procedures and represent a powerful suite of analytical procedures, applicable to a wide range of biomolecules. The remainder of this volume covers a range of chromatographic procedures whose common feature is that they are based on the interaction of a specific ligand with its target protein or other macromolecule. Some of the contributions to this volume are focused on specific groups of molecules, for example complex oligosaccharides and glycosylated proteins (Chapter 5), nucleotidebinding proteins (Chapter 6), proteins that bind free and chelated metal ions (Chapter 10), and DNA-binding proteins (Chapter 11). Others are more generally applicable to a wide range of macromolecules and describe the use of peptides (Chapter 7), inhibitors (Chapter 8), and antibodies (Chapter 9) as affinity ligands, although the latter approaches still rely on specific interaction between immobilized ligand and target macromolecule. In one or two cases, similar protocols are provided in different chapters.

Whilst these could have been edited with brevity in mind, it was felt that that it was preferable to give the choice of the slightly different style of protocol offered by different contributing authors. There are doubtless many other groups of ligands whose chromatographic use could be described. For example, many of the more recent expression vectors permit the overexpression of proteins which bear a specific affinity tag. Appropriate chromatographic matrices are then available to purify the tagged recombinant protein with high selectivity. As the use of these matrices is already well covered by the manufacturer’s instructions they are accordingly not included. However, it is hoped that this volume is reasonably comprehensive and will provide the necessary information in most experimental situations to enable development of rapid and effective purification protocols.

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