Ebook: Separations Using Aqueous Phase Systems: Applications in Cell Biology and Biotechnology

The use of aqueous two-phase systems for the partitioning of macromolecules, organelles and cells was originally developed by Per-Ake Albertsson in Sweden in the mid-fifties [1-3]. These systems were initially applied to separations of plant organelles and viruses but their use has now extended into most areas of cell biology and biochemistry [4,5]. Since 1979 biennial International Conferences on Partitioning in Aqueous Two-Phase Systems have been held in Los Angeles (1979), Sheffield (1981), Vancouver (1983) and Lund (1985). The 5th conference was held in Oxford from 23-28 August 1987 and was entitled "Advances in Separations Using Aqueous Phase Systems in Cell Biology and Biotechnology". It is the formal presentations from this meeting which comprise this volume. In contrast to earlier books on phase partitioning [4,5] this volume contains, for the first time, worldwide contributions from over sixty partitioners from a variety of scientific disciplines, thereby providing a detailed overview of the widespread application and potential of bioseparations using phase partitioning. Disciplines include Biophysics, Biochemistry, Cell Biology, Microbiology, Biotechnology and Process Engineering, in both academic and commercial establishments. These biennial conferences allow advances in these diverse partitioning fields to be reviewed and compared; they also provide an opportunity for those considering using phase partitioning to obtain information, advice and contacts. Attendance has grown steadily over the years and 140 scientists came to Oxford. The conference consisted of ten symposia on areas of application of partitioning which have been organised as specific chapters in this volume.

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Content:
Front Matter....Pages i-xvii
Front Matter....Pages 1-1
General Aspects of Aqueous Polymer Two-Phase Partition....Pages 3-5
Affinity Partitioning of Enzymes....Pages 7-14
Dye Affinity Partitioning of Serum Proteins....Pages 15-23
Hydrophobic Affinity Partitioning as a Method for Studying Conformational Changes in Proteins....Pages 25-31
Affinity Partition of Enzymes with Dextran-Bound Procion Yellow HE-3G: Effect of Ligand Density....Pages 33-34
Rapid Two-Phase Extraction of the Calvin Cycle Enzymes Aldolase and Phosphoglycerate Kinase....Pages 35-37
Affinity Partitioning of Phosphofructokinase from Rat Erythrocytes in Poly(Ethylene Glycol)-Cibacron Blue/Dextran Two-Phase Systems....Pages 39-40
Studies on Triazine Dye-Enzyme Interaction by Means of Affinity Partitioning....Pages 41-42
Front Matter....Pages 43-43
Separation of Organelles and Membranes by Phase Partitioning — A Short Overview....Pages 45-49
Plasma Membrane Isolation by Phase Partition using a Single Partition Step: A Tool for the in vivo Study of Lipid Transfer to the Plasma Membrane....Pages 51-58
Free-Flow Electrophoresis Resolves Plasma Membrane Vesicles Isolated by Aqueous Two Phase Partition into Two Populations of Different Sidedness....Pages 59-60
Raising Monoclonal Antibodies Against Plasma Membranes from Barley Embryos....Pages 61-62
Subfractionation of Cyanobacterial Thylakoids....Pages 63-64
Front Matter....Pages 65-65
Use of Phase Partitioning in Multidimensional Subcellular Fractionation....Pages 67-80
Comparison of Phase Partition with Alternative Methods for Isolation of Subcellular Structures....Pages 81-94
Analysis of the Domain Structure of Membranes by Fragmentation and Separation....Pages 95-99
Two-Phase Partitioning of Rat Liver Plasma Membranes....Pages 101-107
Plasma Membranes from Hepatomas and Cultured Cells by Aqueous Two-Phase Partition....Pages 109-110
Affinity Partitioning and Centrifugal Countercurrent Distribution of Synaptosomes from Calf Brain Cortex....Pages 111-112
Isolation of Plasma Membrane Vesicles from Fat Cells of Epididymal Fat Pads of the Rat by Aqueous Two-Phase Partition....Pages 113-115
Front Matter....Pages 117-117
Separation and Subfractionation of Cell Populations by Phase Partitioning — An Overview....Pages 119-125
Separation of Chondrocytes in Aqueous Two-Phase Systems....Pages 127-135
Problems of White Cell Separation Techniques in Relation to the Phase Partitioning of Neutrophils....Pages 137-144
T Cell Heterogeneity and Lymphoid Cell Populations in Normal Humans and Rheumatoid Arthritis Probed by Partitioning....Pages 145-148
Surface Characterization of Colloidal Drug Carriers for Drug Targeting by Aqueous Two-Phase Partitioning....Pages 149-155
Partition of Algal Cells of Various Strains in Aqueous Two Phase Systems....Pages 157-164
Antibiotic — Neutrophil Interactions Studied by Phase Partitioning....Pages 165-166
Surface Properties of Klebsiella and E Coli: Role of Capsular Polysaccharide in Protection Against Phagocytosis....Pages 167-168
Fractionation by Countercurrent Distribution (CCD) of Rat Erythrocytes During Animal Development....Pages 169-170
Attempts to Fractionate Rat Bone Marrow Cells by Countercurrent Distribution (CCD) in Dextran/Polyethylene Glycol Two-Phase Systems....Pages 171-173
The Partitioning of Liposomes of Defined Size and Composition....Pages 175-177
Front Matter....Pages 179-180
Second Immunoaffinity Ligands for Cell Separation....Pages 181-181
Cell Separation by Immunoaffinity Partition in Aqueous Polymer Two-Phase Systems....Pages 183-191
New Activated PEG Derivatives for Affinity Partitioning....Pages 193-202
Coupling of PEG to Proteins by Activation with Tresyl Chloride. Applications in Immunoaffinity Cell Partitioning....Pages 203-210
Front Matter....Pages 211-213
An Overview of Theoretical Developments for Protein Partitioning in Aqueous Two-Phase Polymer Systems....Pages 215-215
Protein Transport Processes in the Water-Water Interface in Incompatible Two Phase Systems....Pages 217-227
Water: The Third Component in Polymer Two-Phase Systems....Pages 229-232
Contact Angles as an Analytical Tool for Investigating Two-Phase Interactions with Biological Surfaces: A Review....Pages 233-238
Front Matter....Pages 239-247
Macrophage Surface Affinity Changes During Phagocytosis....Pages 215-215
A Polymer Mixing Approach to Chemoattractant-Stimulated Neutrophil Adhesion....Pages 249-259
Mechanism of Cell Partitioning; Equilibrium Contact Angle Measurements Predict Relative, Kinetic Partitioning Behaviour....Pages 261-270
Demixing Kinetics of Phase Separated Polymer Solutions in Microgravity....Pages 271-280
Dextran Coated Glass Slides as a Model for Cell Surface Interactions with Aqueous Two Phase Polymer Systems....Pages 281-286
A Thermodynamic Study of the Binding of the E. coli F41 Adhesin to its Receptor, Human Glycophorin....Pages 287-288
Temperature Dependence of the Phase Equilibria in the PEG-Dextran System — A Theoretical and Experimental Study....Pages 289-290
Front Matter....Pages 291-292
Large Scale Protein Recovery using Aqueous Phase Systems....Pages 293-293
Continuous Crosscurrent Extraction of Proteins in Process Scale....Pages 295-297
Aqueous Two-Phase Partition in Biochemical Recovery from Mammalian Cell Culture....Pages 299-307
Production of ?-Galactosidase Fused Protein A and the Purification of it in an Aqueous Two-Phase System....Pages 309-315
Continuous Affinity Partitioning in a Poly(Ethylene Glycol)-Reppal Pes Aqueous Two-Phase System....Pages 317-324
Fast Downstream Processing of Proteins by Flocculation and Liquid Extraction....Pages 325-331
Gravity Settling of APS — Characteristics and Process Applications....Pages 333-335
Improved Economics of Extractive Enzyme Recovery by Recycling: A Case Study....Pages 337-339
The use of PEG-NADH/Na2Co3 Aqueous Two-Phase Systems for the Large-Scale Preparation of Technical Grade PEG-(Mr 20,000)-N6-(2-Aminoethyl)-NADH....Pages 341-343
Front Matter....Pages 345-347
Extractive Bioconversions in Aqueous Phase Systems....Pages 349-349
Process Simulation of Continuous Bioconversions in Aqueous Two-Phase Systems....Pages 351-360
Basic Trials on Extractive Bioconversion of Glucose to Lactic Acid....Pages 361-369
Integration of Bioconversion and Down-Stream Processing — Starch Hydrolysis in an Aqueous Two-Phase System....Pages 371-375
Front Matter....Pages 377-378
Liquid-Liquid Partition Chromatography of Biopolymers in Aqueous Two-Phase Polymer Systems....Pages 379-379
Application of Liquid-Liquid Partition Chromatography (LLPC) in the Preparation of Steroid Binding Proteins....Pages 381-392
Column Based Liquid/Liquid Separation of Cells Using Aqueous Polymeric Two-Phase Systems....Pages 393-399
A New Approach to Countercurrent Distribution Combining Separation with Analysis in the Biomek Automated Laboratory Workstation....Pages 401-406
Elution Countercurrent Distribution....Pages 407-415
Gravimetric Analysis of Phase Systems using a Microwave Oven and the Rapid Determination of Phase Diagrams....Pages 417-419
Rapid Partitioning of Human Neutrophils by CCD....Pages 421-422
Quantitation of the Resolving Power of CCD using Human Erythrocytes....Pages 423-424
Front Matter....Pages 425-426
New Polymers for Aqueous Two-Phase Systems....Pages 427-427
Protein Partitioning Between Microemulsions and Conjugate Aqueous Phases....Pages 429-438
Immiscible Liquid Affinity Supports....Pages 439-446
Magnetically Enhanced Aqueous Two-Phase Separation....Pages 447-453
Application of Partition Technology to Particle Electrophoresis....Pages 455-461
Formation of Beads in Aqueous Two-Phase Systems....Pages 463-470
Comparison of Cell and Macromolecular Partition in PEG 8000/Dextran T500, Aquaphase PPT or Reppal PES 200 Phase Systems....Pages 471-472
Some Rheological Properties of Perfluorocarbon Emulsions and Their Potential use in Countercurrent Chromatography Distribution and Chromatography Separation Systems....Pages 473-476
Back Matter....Pages 477-478
....Pages 479-504