Protein Liquid Chromatography is a handbook-style guide to liquid chromatography as a tool for isolating and purifying proteins, consisting of 25 individual chapters divided into three parts: Part A covers commonly-used, classic modes of chromatography such as ion-exchange, size-exclusion, and reversed-phase; Part B deals with various target protein classes such as membrane proteins, recombinant proteins, and glycoproteins; and Part C looks at various miscellaneous related topics, including coupling reaction, buffer solution additives, and software. The text as a whole can be viewed as a systematic survey of available methods and how best to use them, but also attempts to provide an exhaustive coverage of each facet. How to solve a specific problem using a chosen method is the overall essence of the volume. The principle philosophy of this compilation is that practical application is everything; therefore, both classical and modern methods are presented in detail, with examples involving conventional, medium- and high-pressure techniques. Over-exposure to history, concept, and theory has deliberately been avoided. The reader will find a wealth of tips and tricks from users for users, including advice on the advantages and disadvantages of each method. Easy-to-read sections on "Getting started now" and "Where to go from here" attempt to provide hands-on, fool-proof detailed practical procedures with complete and even standard model runs for any scientist or technician at work in this area.
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Individual Chromatographic Modes. Ion exchange chromatography (P.H. Roos). Size exclusion chromatography (M. Rögner). Reversed-phase chromatography (H. Schlüter). Hydrophobic interaction chromatography (L.R. Jacob). Hydroxyapatite chromatography (R. Lukačin, W.R. Deppert). Immobilized metal ion affinity chromatography (M. Kastner). Chromatofocusing (R. Lukačin, W.R. Deppert). Dye-ligand affinity chromatography (J. Kirchberger, H.-J. Böhme). Immobilized artificial membrane chromatography (K. Hauer et al.). Liquid-liquid partition chromatography (U.-B. Hansson, C. Wingren). Displacement chromatography (H. Schlüter, J. Jankowski).
Various Target Protein Classes. Integral membrane proteins (G. Welling, S. Welling-Wester). Recombinant proteins (O. Huber, M. Huber-Wunderlich). DNA-binding proteins (M. Velleman). Lectins and glycoconjugates (H.-J. Gabius). Covalent chromatography (D. Whitney). Cell affinity chromatography (D.D. Putnam et al.).
Miscellaneous Chromatographic Aspects. Coupling reactions (E. Müller). Immobilization of nucleotides (E. Müller, M. Morr). Miscellaneous biospecific affinity gels (A.M. Williams). Scale-up of downstream processing (J. Walter). Chromatography on porous matrices (J. Kruip). Chromatography using Strep-tag affinity peptides (H. N. Müller, T.G.M. Schmidt). Buffers and additives (W.R. Deppert, R. Lukačin). Chromatography software (T. Steger-Hartmann).
Supplement. Abbreviations used in this book. Directory of manufacturers and suppliers. Subject index.