Rajni Hatti-Kaul and her expert coauthors combine theory, methodology, and applications in a practical collection of easily reproducible protocols for bioseparations in aqueous two-phase systems (ATPS). The protocols range from established methods to cutting-edge techniques with potential biotechnological applications. Among the methods detailed are those for ATPS preparation and characterization, for partitioning applied to soluble molecules and particulates (including whole cells, membranes, and organelles), and for the isolation and purification of proteins-including a glimpse of large-scale handling of two-phase separations. Practical and informative, with its detailed guidelines allowing researchers to adapt specific systems to their own separation needs, Aqueous Two-Phase Systems: Methods and Protocols demonstrates the scope and utility of two-phase aqueous systems in both basic and applied research.

Preface

v

List of Contributors

xi

Aqueous Two-Phase Systems: A General Overview

1

(10)

Rajni Hatti-Kaul

Part I. Preparation and Characterization of Aqueous Two-Phase Systems

The Phase Diagram

11

(12)

Anita Kaul

Preparation of Aqueous Two-Phase Systems

23

(12)

Daniel Forciniti

Measurement of Some Physical Properties of Aqueous Two-Phase Systems

35

(12)

Donald E. Brooks

Raymond Norris-Jones

Part II. Partitioning Techniques

Single-Step Partitioning in Aqueous Two-Phase Systems

47

(8)

Telma Teixeira Franco

Rajni Hatti-Kaul

Partitio by Countercurrent Distribution (CCD)

55

(10)

Hans-Erik Akerlund

Liquid-Liquid Partition Chromatography (LLPC)

65

(12)

Ulla-Britt Hansson

Christer Wingren

Part III. Partitioning of Soluble Molecules and Particulates

Metal lon Separations in Aqueous Biphasic Systems and with ABEC™ Resins

77

(18)

Jonathan G. Huddleston

Scott T. Griffin

Jianhua Zhang

Heather D. Willauer

Robin D. Rogers

Partition of Amino Acids and Peptides in Aqueous Two-Phase Systems

95

(12)

I-Ming Chu

Wen-Yi Chen

Predicting Partition Coefficients of Small Solutes Based on Hydrophobicity

107

(12)

Mark A. Eiteman

Eukaryotic Cell Partition: Experimental Considerations

119

(24)

James M. Van Alstine

Concentration and Purification of Viruses

143

(16)

Lena Hammar

Isolation of Plant Plasma Membranes and Production of Inside-Out Vesicles

159

(8)

Christer Larsson

Susanne Widell

Isolation of Inside-Out Thylakoid Vesicles

167

(10)

Hans-Erik Akerlund

Two-Phase Partitioning as a Method for Isolation of Tight Plasma Membrane Vesicles from Saccharomyces cerevisiae and from Chlamydomonas reinhardtii

177

(8)

Birgitta Norling

Purification of Cyanobacterial Thylakoid, Plasma, and Outer Membranes by Two-Phase Partitioning

185

(8)

Birgitta Norling

Purification of Plasma Membranes by Affinity Partitioning

193

(8)

Bengt Jergil

Lars Ekblad

Studying the Influence of Salts on Partitioning of Proteins: Isoelectric Point Determination

201

(8)

Daniel Forciniti

Partitioning of chemically Modified Proteins: A Method for Detection of the Resolution of Aqueous Two-Phase Systems

209

(10)

Telma Teixeira Franco

Detection and Analysis of Interactions by Two-Phase Partition

219

(10)

Lars Backman

Cryopartitioning in Two-Phase Systems Containing Organic Solvents

229

(6)

Gote Johansson

Part IV. Isolation and Purification of Proteins by Two-Phase Extraction

Optimization of Extractions in Aqueous Two-Phase Systems

235

(16)

Ulrich Menge

Two-Phase Extraction of Proteins from Cell Debris

251

(8)

Kristina Kohler Van Alstine

Andres Veide

Aqueous Two-Phase Extraction of Proteins from Animal Tissue

259

(12)

Michael J. Boland

Chaotropic Aqueous Two-Phase Systems: Preparation and Uses

271

(10)

Daniel Forciniti

Temperature-Induced Phase Partitioning for Protein Purification

281

(10)

Anita Kaul

Josefine Persson

Folke Tjerneld

Extraction of Amphiphilic Proteins Using Detergent-Based Aqueous Two-Phase Systems

291

(12)

Torsten Minuth

Use of Charged PEG and Dextran Derivatives for Biomolecule Partitioning

303

(12)

Gote Johansson

Affinity Partitioning Using Poly(ethylene glycol) with Covalently Coupled Hydrophobic Groups

315

(14)

Vithaldas P. Shanbhag

Poul Erik H. Jensen

Dye-Ligand Affinity Partitioning of Proteins

329

(16)

Gerhard Kopperschlager

Jurgen Kirchberger

Metal Affinity Protein Partitioning

345

(10)

Roberto Z. Guzman

Javier E. Garcia

Recovery of Proteins and Phase Components

355

(8)

Gote Johansson

Combination of Extraction with Adsorption for Protein Purification

363

(8)

Gabriel Raya-Tonetti

Nora I. Perotti

Integration of Extraction with Affinity Precipitation

371

(10)

Masamichi Kamihira

Rajni Hatti-Kaul

Bo Mattiasson

Affinity Partitioning Using Magnetic Two-Phase Systems

381

(10)

Masamichi Kamihira

Large-Scale Extraction of Proteins

391

(20)

Teresa Cunha

Raquel Aires-Barros

Part V. Bioconversions in Two-Phase Systems

Extractive Bioconversion in Aqueous Two-Phase Systems

411

(8)

Rajni Hatti-Kaul

Enzyme Reaction in Polymer-Polymer Organic Solvent Two-Phase Systems

419

(8)

Bo Mattiasson

Index

427

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Aqueous Two-Phase Systems

9780896035416

0896035417

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