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.

1: Partitioning of Macromolecules.- 1.1 General Aspects of Aqueous Polymer Two-Phase Partition.- 1.2 Affinity Partitioning of Enzymes.- 1.3 Dye Affinity Partitioning of Serum Proteins.- 1.4 Hydrophobic Affinity Partitioning as a Method for Studying Conformational Changes in Proteins.- 1.5 Affinity Partition of Enzymes with Dextran-Bound Procion Yellow HE-3G: Effect of Ligand Density.- 1.6 Rapid Two-Phase Extraction of the Calvin Cycle Enzymes Aldolase and Phosphoglycerate Kinase.- 1.7 Affinity Partitioning of Phosphofructokinase from Rat Erythrocytes in Poly(Ethylene Glycol)-Cibacron Blue/Dextran Two-Phase Systems.- 1.8 Studies on Triazine Dye-Enzyme Interaction by Means of Affinity Partitioning.- 2: Partitioning of Plant Membranes and Organelles.- 2.1 Separation of Organelles and Membranes by Phase Partitioning — A Short Overview.- 2.2 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.- 2.3 Free-Flow Electrophoresis Resolves Plasma Membrane Vesicles Isolated by Aqueous Two Phase Partition into Two Populations of Different Sidedness.- 2.4 Raising Monoclonal Antibodies Against Plasma Membranes from Barley Embryos.- 2.5 Subfractionation of Cyanobacterial Thylakoids.- 3: Partitioning of Animal Membranes and Organelles.- 3.1 Use of Phase Partitioning in Multidimensional Subcellular Fractionation.- 3.2 Comparison of Phase Partition with Alternative Methods for Isolation of Subcellular Structures.- 3.3 Analysis of the Domain Structure of Membranes by Fragmentation and Separation.- 3.4 Two-Phase Partitioning of Rat Liver Plasma Membranes.- 3.5 Plasma Membranes from Hepatomas and Cultured Cells by Aqueous Two-Phase Partition.- 3.6 Affinity Partitioning and Centrifugal Countercurrent Distribution of Synaptosomes from Calf Brain Cortex.- 3.7 Isolation of Plasma Membrane Vesicles from Fat Cells of Epididymal Fat Pads of the Rat by Aqueous Two-Phase Partition.- 4: Partitioning of Cells.- 4.1 Separation and Subfractionation of Cell Populations by Phase Partitioning — an Overview.- 4.2 Separation of Chondrocytes in Aqueous Two-Phase Systems.- 4.3 Problems of White Cell Separation Techniques in Relation to the Phase Partitioning of Neutrophils.- 4.4 Use of Partitioning to Study Differentiating Cells of Dictyostelium Discoideum.- 4.5 Surface Characterization of Colloidal Drug Carriers for Drug Targeting by Aqueous Two-Phase Partitioning.- 4.6 Use of Partitioning to Study Differentiating Cells of Dictyostelium Discoideum.- 4.7 Partition of Algal Cells of Various Strains in Aqueous Two Phase Systems.- 4.8 Antibiotic — Neutrophil Interactions Studied by Phase Partitioning.- 4.9 Surface Properties of Klebsiella and E Coli: Role of Capsular Polysaccharide in Protection Against Phagocytosis.- 4.10 Fractionation by Countercurrent Distribution (CCD) of Rat Erythrocytes During Animal Development.- 4.11 Attempts to Fractionate Rat Bone Marrow Cells by Countercurrent Distribution (CCD) in Dextran/Polyethylene Glycol Two-Phase Systems.- 4.12 The Partitioning of Liposomes of Defined Size and Composition.- 5: Immuno-Affinity Partitioning.- 5.1 Second Immunoaffinity Ligands for Cell Separation.- 5.2 Cell Separation by Immunoaffinity Partition in Aqueous Polymer Two-Phase Systems.- 5.3 New Activated PEG Derivatives for Affinity Partitioning.- 5.4 Coupling of PEG to Proteins by Activation with Tresyl Chloride. Applications in Immunoaffinity Cell Partitioning.- 6: Theoretical Aspects of Phase Partitioning.- 6.1 An Overview of Theoretical Developments for Protein Partitioning in Aqueous Two-Phase Polymer Systems.- 6.2 Protein Transport Processes in the Water-Water Interface in Incompatible Two Phase Systems.- 6.3 Water: The Third Component in Polymer Two-Phase Systems.- 6.4 Contact Angles as an Analytical Tool for Investigating Two-Phase Interactions with Biological Surfaces: A Review.- 6.5 Macrophage Surface Affinity Changes During Phagocytosis.- 6.6 A Polymer Mixing Approach to Chemoattractant-Stimulated Neutrophil Adhesion.- 6.7 Mechanism of Cell Partitioning; Equilibrium Contact Angle Measurements Predict Relative, Kinetic Partitioning Behaviour.- 6.8 Demixing Kinetics of Phase Separated Polymer Solutions in Microgravity.- 6.9 Dextran Coated Glass Slides as a Model for Cell Surface Interactions with Aqueous Two-Phase Polymer Systems.- 6.10 A Thermodynamic Study of the Binding of the E. coli F41 Adhesin to its Receptor, Human Glycophorin.- 6.11 Temperature Dependence of the Phase Equilibria in the PEG-Dextran System — A Theoretical and Experimental Study.- 7: Biotechnology: Downstream Processing.- 7.1 Large Scale Protein Recovery using Aqueous Phase Systems.- 7.2 Continuous Crosscurrent Extraction of Proteins in Process Scale.- 7.3 Aqueous Two-Phase Partition in Biochemical Recovery from Mammalian Cell Culture.- 7.4 Production of ß-Galactosidase Fused Protein a and the Purification of it in an Aqueous Two-Phase System.- 7.5 Continuous Affinity Partitioning in a Poly-(Ethylene Glycol)-Reppal Pes Aqueous Two-Phase System.- 7.6 Fast Downstream Processing of Proteins by Flocculation and Liquid Extraction.- 7.7 Gravity Settling of APS — Characteristics and Process Applications.- 7.8 Improved Economics of Extractive Enzyme Recovery by Recycling: A Case Study.- 7.9 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.- 8: Biotechnology: Extractive Bioconversions.- 8.1 Extractive Bioconversions in Aqueous Phase Systems.- 8.2 Process Simulation of Continuous Bioconversions in Aqueous Two-Phase Systems.- 8.3 Basic Trials on Extractive Bioconversion of Glucose to Lactic Acid.- 8.4 Integration of Bioconversion and Down-Stream Processing — Starch Hydrolysis in an Aqueous Two-Phase System.- 9: Emerging Technology: New Methodology Involving Liquid-Liquid Chromatography and Countercurrent Distribution.- 9.1 Liquid-Liquid Partition Chromatography of Biopolymers in Aqueous Two-Phase Polymer Systems.- 9.2 Application of Liquid-Liquid Partition Chromatography (LLPC) in the Preparation of Steroid Binding Proteins.- 9.3 Column Based Liquid/Liquid Separation of Cells using Aqueous Polymeric Two-Phase Systems.- 9.4 A New Approach to Countercurrent Distribution Combining Separation with Analysis in the Biomek Automated Laboratory Workstation.- 9.5 Elution Countercurrent Distribution.- 9.6 Gravimetric Analysis of Phase Systems using a Microwave Oven and the Rapid Determination of Phase Diagrams.- 9.7 Rapid Partitioning of Human Neutrophils by CCD.- 9.8 Quantitation of the Resolving Power of CCD using Human Erythrocytes.- 10: Emerging Technology: New Polymers and Phase Systems.- 10.1 New Polymers for Aqueous Two-Phase Systems.- 10.2 Protein Partitioning Between Microemulsions and Conjugate Aqueous Phases.- 10.3 Immiscible Liquid Affinity Supports.- 10.4 Magnetically Enhanced Aqueous Two-Phase Separation.- 10.5 Application of Partition Technology to Particle Electrophoresis.- 10.6 Formation of Beads in Aqueous Two-Phase Systems.- 10.7 Comparison of Cell and Macromolecular Partition in PEG 8000/Dextran T500, Aquaphase PPT or Reppal PES 200 Phase Systems.- 10.8 Some Rheological Properties of Perfluorocarbon Emulsions and Their Potential use in Countercurrent Chromatography Distribution and Chromatography Separation Systems.