Column Handbook for Size Exclusion Chromatography

Front Cover......Page 1
Column Handbook for Size Exclusion Chromatography......Page 4
Copyright Page......Page 5
CONTENTS......Page 6
CONTRIBUTORS......Page 20
PREFACE......Page 22
PART I: CONTRIBUTIONS FROM SIZE EXCLUSION CHROMATOGRAPHY AND GEL-FILTRATION CHROMATOGRAPHY COLUMN MANUFACTURERS......Page 26
I. Introduction......Page 28
II. Basic Features of Conventional Suspension Polymerization......Page 29
III. Preparation of Organic Polymeric Beads by Conventional Suspension Polymerization......Page 32
IV. Special Polymeric Supports......Page 35
V. Basic Features of Monosized Polymeric Particles Prepared by Activated Swelling and Polymerization......Page 40
VI. Application of Polymeric Beads as Size Exclusion Chromatography Supports......Page 44
References......Page 49
I. Introduction: Development of Size Exclusion Chromatography (SEC)......Page 52
II. Application Areas of SEC......Page 54
III. Characteristics of Gel-Filtration/Size Exclusion Media......Page 64
IV. Columns for Packing SEC Media......Page 79
V. Preparation of the Size Exclusion Column......Page 86
VI. Optimization of Gel-Filtration/Size Exclusion Separations......Page 92
References......Page 98
I. Background Information......Page 100
II. Using and Selecting Zorbax PSM Columns and Mobile Phases......Page 103
III. HPSEC Troubleshooting......Page 107
IV. Applications......Page 110
References......Page 117
I. Silica-Based TSK-GEL SW/SWXL Columns for Gel-Filtration Chromatography (GFC)......Page 118
II. Polymer-Based TSK-GEL PW/PWXL Columns for Aqueous GFC......Page 131
III. Choosing TSK-GEL SW Type or TSK-GEL PW Type Columns for GFC......Page 153
IV. Cleaning and Rehydration of TSK-GEL Columns......Page 159
V. TSK-GEL H Type Columns......Page 160
VI. Toyopearl Resins for Size Exclusion Chromatography......Page 170
References......Page 182
I. Introduction......Page 184
II. American Polymer Gel-Permeation/Size Exclusion Chromatography (GPC/SEC) Column Repacking......Page 185
IV. Manufacture of GPC/SEC Gels......Page 188
V. Conclusion......Page 194
II. Gel Technology of Shodex Columns......Page 196
III. Shodex Organic GPC Series......Page 197
IV. Shodex OHpak SB-800HQ Series......Page 218
V. Shodex Asahipak GS/GF Series......Page 222
VI. Shodex Protein KW Series......Page 229
VII. Conclusion......Page 235
I. Introduction to Size Exclusion Chromatography (SEC)......Page 244
II. Synthesis of Fractogel EMD BioSEC......Page 245
III. Properties of Fractogel EMD BioSEC......Page 246
IV. Adequate Buffers......Page 247
V. Recommended Columns for SEC......Page 249
VI. Packing the Column......Page 251
VII. Recommended Flow Rates for SEC Columns......Page 257
VIII. Loading Capacity......Page 258
IX. Determination of Molecular Mass of Proteins......Page 260
X. Column and Gel Maintenance, Cleaning, Regeneration, and Storage Methods......Page 262
XI. Application Areas......Page 266
XII. Troubleshooting......Page 271
References......Page 272
I. Overview of Size Exclusion Chromatography (SEC) of Small Solutes......Page 274
II. Introduction of PolyHYDROXYETHYL Aspartamide and the Effect of Chaotropes on the Fractionation Range......Page 275
III. Verification of SEC Mechanism for Small Solutes......Page 279
IV. Fractionation Range with PolyHEA as a Function of Pore Diameter......Page 285
References......Page 290
I. Introduction......Page 292
II. Basis of SEC Column Design......Page 293
III. Review: SEC Sorbents and Requirements......Page 295
IV. SEC Column Selection Criteria......Page 296
V. Testing and Evaluation of SEC Columns......Page 308
VI. PSS Column Quality Control System......Page 312
VII. Applications of PSS SEC Columns......Page 316
VIII. Conclusion......Page 323
References......Page 328
I. Introduction......Page 330
II. Support Technology......Page 331
III. Column Technology......Page 335
IV. Operation......Page 339
References......Page 348
I. Introduction......Page 350
II. Column Selection Principles......Page 351
III. Waters Products for Size Exclusion Chromatography......Page 360
IV. Summary......Page 372
References......Page 373
I. Introduction......Page 374
II. PLgel Organic Size Exclusion Chromatography (SEC) Columns......Page 375
III. PL HFIPgel Columns......Page 384
IV. PL aquagel-OH Aqueous SEC Columns......Page 385
V. Column Hardware......Page 388
References......Page 391
I. Introduction......Page 392
II. The Basic Polydivinylbenzene Packing......Page 393
III. Basic and Chemically Modified Polydivinylbenzene Packings......Page 397
IV. Modified GBR/PDVB Gel......Page 401
V. Discussion of Chromatographic Applications of Various Bonded and Nonbonded Phases......Page 404
References......Page 450
PART II: CONTRIBUTIONS FROM USERS OF SIZE EXCLUSION CHROMATOGRAPHY AND GEL-PERMEATION CHROMATOGRAPHY......Page 452
I. Introduction......Page 454
II. Determination of the Number of Theoretical Plates......Page 456
IV. Separation Efficiency......Page 460
V. Apparent Pore Size Distribution......Page 462
VI. Surface Interactions......Page 463
VII. Separation of the Oligomer Range from System Peaks......Page 464
VIII. Column Dimensions......Page 465
IX. Conclusion......Page 466
References......Page 468
I. Introduction......Page 470
II. Experimental......Page 473
III. Results......Page 475
IV. Discussion......Page 480
References......Page 482
I. Introduction......Page 484
II. Dextran Gels......Page 490
III. Combined System: Fractogel TSK HW40 (s)/HW50 (s)/Superose 6 (Prep Grade)......Page 502
IV. Agarose Gels......Page 504
V. Combined System: Bio-Gel P-6/Sephacryl S-200......Page 509
VI. Prepacked System: TSK-GEL PW Columns......Page 514
References......Page 522
I. Introduction......Page 524
II. Experimental......Page 526
III. Results and Discussion......Page 527
IV. Conclusions......Page 549
References......Page 554
II. Theory and Primary Applications......Page 556
III. Examples......Page 557
IV. Assay Development and Validation......Page 559
V. Specialized Applications......Page 560
References......Page 561
I. Introduction......Page 564
II. Solubility and Adsorption......Page 566
III. General GPC Practices......Page 569
IV. Nonroutine Uses of GPC Columns......Page 576
V. GPC of Acrylates Using Solvents Other Than Tetrahydrofuran......Page 578
VI. Polyacrylic Acid and Related Polymers......Page 579
References......Page 582
I. Introduction......Page 584
II. Aqueous SEC Using TSK-PW Columns......Page 586
III. Aqueous SEC Using Synchropak Columns......Page 596
IV. Aqueous SEC Using Cationic Columns......Page 598
V. Summary......Page 604
References......Page 605
I. Introduction......Page 608
III. Column Selection Criteria......Page 609
IV. Quality Control Procedure......Page 614
References......Page 617
PART III:  COLUMNS FOR OTHER RELATED POLYMER SEPARATION OR FRACTIONATION TECHNIQUES......Page 620
II. Open Microcapillary Tubular Hydrodynamic Chromatography (OTHdC)......Page 622
III. Packed Column HdC (PCHdC)......Page 626
IV. Comparison to Other Separation Techniques......Page 632
References......Page 635
I. What Is High Osmotic Pressure Chromatography (HOPC)?......Page 636
II. Separation Principle......Page 639
III. Packing Materials......Page 642
IV. Pore Size and Particle Size......Page 647
V. Column Dimensions......Page 651
VI. Summary......Page 653
References......Page 654
INDEX......Page 656