Partitioning In Aqueous Two - Phase System

Partitioning in Aqueous Two-Phase Systems: Theory, Methods, Uses, and Applications to Biotechnology is a collection of papers that discusses the applications of aqueous two-phase systems to problems of separation and extraction of macromolecules, organelles, and cells. Papers focus on the theoretical basis and the practical details of the procedures used. Some of the papers describe in one or a few steps how two components can be separated by the investigator manipulating their partitions so that one component is in one phase and the other component is in the other phase or at the interface. Investigators can also avail of developed batch extractions for plant organelles, cell membranes, nucleic acids, and proteins. The book cites as an example the partitioning of right-side-out and inside-out vesicles (obtained from fragments of thylakoid membranes) to the top and bottom phases, respectively, of a Dx-PEG system. Other papers describe the use of the countercurrent distribution when single extraction steps are not sufficient to produce a separation in materials that do not differ greatly in their partitioning behavior. The collection can prove valuable for bio-chemists, cellular biologists, micro-biologists, and developmental biologists.

Contenu

Contributors
Preface
Abbreviations and Conventions
1 History of Aqueous Polymer Two-Phase Partition
Text
References
2 Theoretical Aspects of Partitioning
I. Properties of Polymers
II. Phase Separation
III. Theory of Partitioning
References
3 Preparation of Phase Systems and Measurement of Their Physicochemical Properties
I. Introduction
II. Preparation of Phase Systems
III. Single-Step Partition Experiments
IV. Measurement of Physicochemical Properties of Phase Systems
References
4 Thin-Layer Countercurrent Distribution and Apparatus
I. Introduction
II. Principles of Countercurrent Distribution
III. Phase System Selection
IV. Countercurrent Distribution of Particulate Samples
V. Thin-Layer Countercurrent Distribution Apparatus
References
5 Other Types of Countercurrent Distribution Apparatus and Continuous Flow Chromatography Techniques
I. Introduction
II. Enhanced Gravity Countercurrent Distribution
III. Column Chromatography
IV. Countercurrent Chromatography
V. Emerging Techniques
VI. Commercial Availability
References
6 Partitioning of Proteins
I. Introduction
II. Properties Determining the Partitioning Behavior of Proteins
III. Practical Procedures Used in Partitioning Proteins
IV. Practical Aspects of Protein Extraction
V. Multistep Extraction Procedures
VI. Analytical Applications
VII. Specific Studies
References
7 Partitioning of Nucleic Acids
I. Introductory Remarks
II. General Behavior of Nucleic Acids in Aqueous Two-Phase Systems
III. Isolation and Fractionation of Nucleic Acids in Aqueous Two-Phase Polymer Systems
IV. Fractionation of Chromosomes by Partitioning in Poly(ethylene Glycol)-Dextran Systems
V. Applications of Partitioning for Studying Nucleic Acid-Ligand Interactions
References
8 Interacting Systems and Binding Studied by Partitioning
I. Introduction
II. Theory
III. Examples
IV. Conclusion
References
9 Steroid Receptors and Steroid-Binding Plasma Proteins Studied by Partitioning
I. Biological Functions of Steroid-Binding Proteins
II. Use of Partitioning for Studying Steroid-Protein Binding
III. Use of Partitioning for Studying Molecular Conversions of Glucocorticoid-Receptor Complexes
References
10 Surface Properties of Cells Reflected by Partitioning: Red Blood Cells as a Model
I. Introduction
II. What Red Blood Cells Taught Us About Phase Systems
III. What Phase Systems Taught Us About Red Blood Cells
IV. Comments on Dextran-Ficoll Aqueous Phase Systems
V. Conclusion
References
11 Separation and Subfractionation of Selected Mammalian Cell Populations
I. Introduction
II. Separation and Subfractionation of Blood Elements
III. Partitioning of Bone Marrow Cells: Cells Containing Hemoglobin
IV. Fractionation of Lymphoid Cells from Spleen and Other Tissues
V. Tissue Cells
VI. Tissue Culture Cells
VII. Conclusion
References
12 Partitioning of Bacteria, Virus, and Phage
I. Introduction
II. Aqueous Two-Phase Partitioning of Bacteria
III. Aqueous Two-Phase Partitioning of Virus and Phage
IV. Summary
References
13 Partitioning of Animal Membranes and Organelles
I. Introduction
II. Relevant Principles in Phase Partitioning as Applied to Cell Particles
III. Preparative and Semipreparative Procedures
IV. Analytical Phase Partitioning Procedures: Application to Membrane Fragments and Organelles
V. Conclusions
References
14 Partitioning of Plant Cells, Cell Walls, Membranes, and Organelles
I. Cells and Cell Walls
II. Protoplasts
III. Plasma Membranes
IV. Intact Chloroplasts
V. Leaf Mitochondria
VI. Separation of Right-Side-Out and Inside-Out Membrane Vesicles
VII. Concluding Remarks
References
15 Applications of Phase Partitioning in Biotechnology
I. Introduction
II. Enzyme Purification by Liquid-Liquid Extraction in Aqueous Two-Phase Systems
III. Extractive Bioconversions in Aqueous Two-Phase Systems
IV. Additional Preparative Applications
Appendix: Analysis of PEG-Salt Systems
References
16 Polymer-Ligands Used in Affinity Partitioning and Their Synthesis
I. Introduction
II. General Chemistry of Poly(ethylene Glycol)
III. Synthesis of Poly(ethylene Glycol) Derivatives
IV. Selected Poly(ethylene Glycol) Derivatives for Affinity Partitioning
V. General Chemistry of Dextran
VI. Synthesis of Dextran Derivatives
VII. Selected Dextran Derivatives for Affinity Partitioning
References
17 Partitioning: A Comprehensive Bibliography
Text
Bibliography
Addendum 1961-1983
Addendum 1984-1985
Index