Protein purification : priciples and practice

목차 (Table of Contents)

• CONTENTS
• Series Preface = ⅴ
• Preface to the Third Edition = ⅶ
• Preface to the Second Edition = ⅸ
• Preface to the First Edition = xi
• Chapter 1 The Protein Purification Laboratory = 1
• 1.1 Apparatus, Special Materials, and Reagents = 1
• 1.2 Separation of Precipitates and Particulate Material = 3
• Filtration = 3
• Centrifugation = 4
• 1.3 Principles of Column Chromatography = 8
• 1.4 Manipulation of Protein Solutions = 14
• Concentration = 15
• Removal of Salts； Changing Buffers = 17
• Chapter 2 Making an Extract = 22
• 2.1 The Raw Material = 22
• Freshness and Storage = 25
• 2.2 Cell Disintegration and Extraction = 26
• Mammalian Tissues = 30
• Erythrocytes = 31
• Soft Plant Tissues = 31
• Yeasts = 31
• Bacteria = 32
• Fatty Tissues = 34
• 2.3 Optimization and Clarification of the Extract = 34
• 2.4 Extraction of Membrane Proteins = 38
• Chapter 3 Analysis―Measurement of Protein and Enzyme Activity = 44
• 3.1 Methods for Measuring Protein Concentration = 44
• Biuret Reaction = 45
• Lowry Method = 46
• UV Absorption = 46
• Dye Binding = 48
• Bicinchonic Acid = 48
• 3.2 Measurement of Enzyme Activity―Basic Principles = 50
• Substrate Concentration, Activators, and Inhibitors = 50
• pH, Ionic Strength, and Temperature = 55
• 3.3 Measurement of Enzyme Activity Using Stopped Methods = 56
• Incubation Conditions = 57
• Stopping Methods = 58
• Measurement of Product = 59
• 3.4 Measurement of Enzyme Activity Using Continuous Methods = 62
• Coupled Methods = 63
• 3.5 Practical Points in Enzyme Activity Determination = 68
• Chapter 4 Separation by Precipitation = 71
• 4.1 General Observations = 71
• 4.2 The Solubility of Proteins at Low Salt Concentrations = 72
• Points to Note in Practice = 75
• 4.3 Salting Out at High Salt Concentration = 76
• 4.4 Precipitation with Organic Solvents = 85
• General Theory = 85
• Choice ot Solvent = 87
• Operating Procedures = 89
• 4.5 Precipitation with Organic Polymers and Other Materials = 92
• 4.6 Affinity Precipitation = 93
• 4.7 Precipitation by Selective Denaturation = 95
• General Principles = 95
• Temperature Denaturation = 96
• pH Denaturation = 98
• Denaturation by Organic Solvents = 100
• Chapter 5 Separation by Adsorption I: General Principles = 102
• 5.1 General Chromatographic Theory = 103
• Partition Coefficients = 103
• Zone Spreading, Resolution, and the Plate Height Concept = 105
• The Dissociation Constant for Protein-Adsorbent Interaction = 111
• Simplified Theory of Adsorption = 112
• 5.2 Membrane Adsorbents； Radial Flow Columns = 119
• 5.3 Batch Adsorption = 121
• General Principles = 121
• Practical Approaches = 123
• 5.4 High-Performance Liquid Chromatography = 126
• General Principles = 126
• Relationships Between Bead Size, Flow Rate, Pressure, and Optimum Performance = 128
• 5.5 Types of Adsorbent Used in Protein Chromatography = 132
• Nature of the Bead Matrix = 132
• Summary of Adsorbent Types = 135
• 5.6 Operating Conditions for Column Chromatography = 139
• Sample Application = 139
• Overload and Displacement Chromatography = 139
• Flow Rates = 142
• Chapter 6 Separation by Adsorption II: Ion Exchangers and Nonspecific Adsorbents = 146
• 6.1 Ion Exchangers―Principles, Properties, and Uses = 146
• General Principles = 146
• Adsorptive Capacities of Ion Exchangers = 150
• Types of Ion Exchangers = 152
• pH and Donnan Effects = 153
• Elution of Adsorbed Protein = 154
• 6.2 Ion-Exchange Chromatograpahy―Practical Aspects = 157
• Trials to Determine Ion-Exchange Behavior = 159
• Buffers for Use in Ion-Exchange Chromatography = 160
• Conditions of Adsorption = 164
• Size and Dimensions of the Column = 165
• Procedures for Elution = 167
• 6.3 Inorganic Adsorbents = 172
• Hydroxyapatite and Calcium Phosphate Gels = 173
• 6.4 Hydrophobic Adsorbents = 175
• Application of Sample to a Hydrophobic Column = 176
• Elution of Protein from Hydrophobic Columns = 177
• Reverse Phase Chromatography = 178
• Other Hydrophobic Techniques = 179
• 6.5 Immobilized Metal Affinity Chromatography (IMAC) = 180
• General Principles = 180
• Operating Conditions for IMAC = 182
• 6.6 Miscellaneous Adsorbents = 183
• Cationic Polymer-Nucleic Acid Complexes as Batch Adsorbents = 183
• Thiophilic Adsorbents = 184
• Mixed-Function Adsorbents = 185
• Chapter 7 Separation by Adsorption―Affinity Techniques = 187
• 7.1 Principles of Affinity Chromatography = 187
• Synthesis of Affinity Adsorbents = 188
• Application of Chromatographic Theory to Affinity Adsorbents = 196
• General Techniques and Procedures in Affinity Adsorption Chromatography = 200
• 7.2 Immunoadsorbents = 204
• Basic Principles = 205
• Methods Using Polyclonal Antibodies = 205
• Methods Using Monoclonal Antibodies = 208
• Relative Advantages of Polyclonal and Monoclonal Antibodies = 209
• 7.3 Dye Ligand Chromatography = 210
• Developmental History = 210
• Preparation of Dye Ligand Adsorbents = 214
• Dye-Protein Interactions = 217
• Screening to Obtain a Suitable Adsorbent = 219
• Elution of Proteins and Enzymes from Dye Columns = 223
• Cleaning and Storage of Dye Adsorbents = 224
• 7.4 Affinity Elution from Ion Exchangers and Other Adsorbents = 226
• Affinity Elution from Ion Exchangers = 227
• Affinity Elution from Other Adsorbents = 231
• Practice and Theory of Affinity Elution = 233
• 7.5 Commonly Used Affinity and Pseudo-Affinity Adsorbents = 236
• Small Ligands = 236
• Biopotymer Ligands = 236
• Chapter 8 Separation in Solution = 238
• 8.1 Gel Filtration = 238
• Practical Procedures = 243
• 8.2 Electrophoretic Methods = 250
• Electrophoresis Principles = 251
• Methods for Preparative Electrophoresis―Horizontal Slabs = 253
• Methods for Preparative Eleclrophoresis―Vertical Systems = 255
• Buffer Systems for Electrophoresis = 256
• Isoetectric Focusing = 258
• Isotachophoresis = 262
• 8.3 Liquid Phase Partitioning = 264
• 8.4 Ultrafiltration = 267
• Chapter 9 Purification of Special Types of Proteins = 270
• 9.1 Recombinant Proteins = 270
• Terminology of Recombinant Proteins = 271
• 9.2 Membrane Proteins = 277
• 9.3 Purification of Antibodies = 279
• Chapter 10 Small-Scale and Large-Scale Procedures = 283
• 10.1 Small-Scale Procedures―Proteins for Sequencing = 283
• 10.2 Large-Scale Procedures = 287
• Scaling Up in the Laboratory = 287
• Commercial-Scale Protein Production = 291
• Chapter 11 Analysis for Purity = 293
• 11.1 Electrophoretic Analysis = 293
• Simple (Native) Gel Electrophoresis = 294
• Urea Gels = 296
• SDS Gels = 296
• Gradieni Gels = 298
• Isoelectric Focusing = 299
• Two-Dimensional Systems = 300
• Capillary Electrophoresis = 300
• Staining and Detection of Proteins after Electrophoresis = 302
• Detection of Specific Proteins = 303
• 11.2 Other Analytical Methods = 307
• Chapter 12 Optimization of Procedures；Final Steps = 310
• 12.1 Speed Versus Resolution: The Time Factor = 311
• 12.2 Stabilizing Factors for Enzymes and Other Proteins = 317
• Prevention of Denaturation = 317
• Avoidance of Catalytic Site Inaclivation = 318
• Avoidance of Proteolytic Degradation = 321
• Other Stabilizing Influences on Proteins = 323
• 12.3 Control of pH: Buffers = 324
• Buffer Theory = 324
• Effect of Temperature, Ionic Strength, and Organic Solvents on pK $$pK_a$$ Values = 326
• Making Up Buffer Solutions = 330
• 12.4 Following a Published Procedure = 333
• 12.5 Final Steps―Storage, Crystallization, and Publication = 335
• Crystallization for Purification = 336
• Methods for Crystallization for X-ray Diffraction Studies = 337
• Conditions for Storage of Purified Proteins = 342
• What is Important for Publication? = 344
• Appendix A Precipitation Tables = 346
• Appendix B Solutions for Measuring Protein Concentration = 349
• Appendix C Buffers for Use in Protein Chemistry = 351
• Appendix D Chromatographic Materials = 353
• References = 356
• Index = 375