Gel Chromatography Gel Filtration - Gel Permeation - Molecular Sieves: A Laboratory Handbook

ISBN-13: 9783642950841 / Angielski / Miękka / 2012 / 196 str.

Helmut Determann; E. Gross; J. M. Harkin

Opis

The efforts spent on many a scientific book cannot be justified, no matter how many words are said about it. The opposite is true for this book and a few brief remarks upon its publication. Within a short period of time, short even by all present standards, gel chromatography has gone through a development and experienced an acceptance that is unknown to any other method. From experience, the new and unique separation technique is today known and liked in all laboratories that are concerned with substances of high molecular weight; in others, the technique is known from hearsay, the least. Soon it became evident that a comprehensive coverage of the conceptual development, the theoretical principles, and the experimental technique of the new method would be desirable. This coverage is now offered by the book of an expert. Its author has personally participated in the development from its beginning and helped to promote it. He has made possible the gel chromatography, also of 'proteins, on thin layer plates; for lipophilic substances he has contributed considerably to the transition from water to organic solvent systems and developed theoretical concepts for a better understanding of the effects that are responsible for the separation. The book, so it appears to me, is pointing in new directions. The reader does not only expect a clear presentation of facts but also that of instructions for practical applications. Both these expectations have been met by the expert.

Spis treści

1 Introduction.- Separation by Molecular Weight Differences.- Dialysis.- Homogeneous Gels.- Chromatography on Granulated Gels.- Illustrative Experiment.- Terminology.- Historical Survey.- Ion Exchangers.- Neutral Gels.- References.- 2 Materials and Methods.- The Gels.- The Preparation of Hydrophylic Gels.- Dextran Gels.- Polyacrylamide Gels.- Agar Gels, Agarose Gels.- The Preparation of Organophilic Gels.- Sephadex Derivatives.- Acrylic Gels.- Polystyrene Gels.- Aerogels.- Commercial Gels.- Sephadex.- Sephadex LH-20.- Bio-Gel.- Agarose.- Styragel.- Equipment.- Columns.- Pumps.- Hose Connections.- Detectors.- Fraction Collectors.- Integrated Units.- Experimental Techniques with Columns.- Packing of the Gel Bed.- Chromatography.- Sample Application.- Elution.- Analysis.- Extension of the Gel Bed.- Recycling Chromatography.- Thinlayer Gel Chromatography.- Preparation.- Chromatography.- Results.- Centrifuge Techniques.- Equipment.- Illustrative Experiments.- Desalting.- Concentrating.- References.- 3 Theory.- Numerical Evaluation of Experimental Data.- Volumes in the Gel Bed.- Elution Parameters.- Effectivity of a Gel Column.- Approaches of Interpretation.- The Exclusion Principle.- Simple Geometric Models.- The Ogston-Laurent Gel Model.- The Principle of Restricted Diffusion.- Comparison of the Various Interpretations.- The Partition Principle.- Affinity of Solutes to the Gel Phase.- Partition and Adsorption.- Hydrophylic Gels.- References.- 4 Principles of Application.- Gel Filtration.- Desalting.- Group Separation.- Modification of Macromolecules.- Complex Formation.- Gel Chromatography.- Molecular Weight Determination.- Calibration.- Technique.- Molecular Weight Determination of Proteins.- Theory.- Results.- Association Equilibria.- The Study of Polydisperse Systems.- General Principles.- Gel Chromatography.- Evaluation of Gel Permeation Chromatograms.- Results.- Separation without Differences in Size.- Chromatography in Aqueous Media.- Chromatography with Organic Solvents.- Fractional Precipitation.- References.- 5 Results.- Enzymology.- Isolation of Enzymes.- Enzymatic Reactions.- Endocrinology.- Chemistry of Plasma Proteins.- Structural Elucidation of Proteins.- Nucleic Acid Chemistry.- Nucleic Acids.- Oligonucleotides.- Virus Separation.- Carbohydrates.- Clinical Chemistry.- Miscellaneous Applications.- References.- Author Index.