Handbook of Bioseparations
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Table of contents
- CONTENTSv
- PREFACExiii
- Chapter 1. Bioseparations: An Overview1
- I. Introduction1
- II. Analytical Methodologies3
- III. Separation and Purification Methods9
- IV. Other Important Considerations18
- Reference21
- Chapter 2. Analysis of Protein Impurities in Pharmaceuticals Derived from Recombinant DNA23
- I. Introduction23
- II. Protein Impurity Analysis28
- III. Experimental Summary57
- IV. Case Studies58
- References64
- Chapter 3. Physicochemical Factors in Polypeptide and Protein Purification and Analysis by High-Perf71
- I. Introduction72
- II. Basic Chromatographic Terms and Concepts76
- III. The Chemical Structure of Polypeptides and Proteins79
- IV. Physicochemical Factors That Underpin Ligate Interactions with Polypeptides and Proteins in HPLC84
- V. Strategic Considerations behind the HPLC Separations107
- VI. Specific Physicochemical Considerations on the Individual Chromatographic Modes117
- VII. The Effect of Temperature and the Thermodynamics of Polypeptide– or Protein–Ligate Interact135
- VIII. Factors That Control Performance and Efficiency156
- IX. Scaling-Up Possibilities: Heuristic Approaches and Productivity Considerations172
- X. Effect of Mass Transfer Resistances in Preparative HPLC of Polypeptides and Proteins178
- XI. Summary218
- References222
- Chapter 4. Capillary Electrophoresis of Compounds of Biological Interest237
- I. Introduction238
- II. Capillary Zone Electrophoresis239
- III. Migration Behavior of Peptides and Proteins243
- IV. Modifications of Fused Silica Capillaries247
- V. Effect of Temperature on Separations252
- VI. Strategy for Protein Separations252
- VII. Capillary Gel Electrophoresis253
- VIII. Micellar Electrokinetic Chromatography255
- IX. Capillary Electrochromatography255
- X. Applications256
- References262
- Chapter 5. Isoelectric Focusing263
- I. Introduction263
- II. The Principles of Isoelectric Focusing264
- III. Analytical Isoelectric Focusing276
- IV. Two-Dimensional Gel Electrophoresis285
- V. Preparative Isoelectric Focusing287
- VI. Capillary Isoelectric Focusing291
- VII. Summary292
- VIII. Appendix292
- References295
- Chapter 6. Mass Spectrometry of Biomolecules299
- I. Introduction299
- II. Examples of Applications of Mass Spectrometry to Biological Research309
- III. Conclusions325
- References325
- Chapter 7. Liquid–Liquid Partitioning Methods for Bioseparations329
- I. Introduction329
- II. Solvent Extraction for Bioseparations330
- III. Aqueous Two-Phase Partitioning for Bioseparations348
- IV. Summary360
- References361
- Chapter 8. Separation of Nucleic Acids and Proteins365
- I. Introduction365
- II. Precipitation of Nucleic Acids368
- III. Nuclease Treatment370
- IV. Aqueous Two-Phase Extraction370
- V. A Case Study372
- VI. Conclusions376
- References376
- Chapter 9. Bioseparations by Displacement Chromatography379
- I. Introduction380
- II. Purification of Amino Acids and Peptides by Displacement Chromatography382
- III. Purification of Proteins by Displacement Chromatography383
- IV. Alternative Modes of Displacement Chromatography390
- V. Methods Development for Displacement Chromatography393
- VI. Displacement Chromatography for the Purification of Biomolecules: Industrial Case Studies400
- VII. Design of Low Molecular Weight Displacers406
- VIII. Conclusions410
- References412
- Chapter 10. Physicochemical Basis of Expanded-Bed Adsorption for Protein Purification417
- I. Introduction417
- II. Typical Procedure to Operate an Expanded-Bed Chromatographic System418
- III. Ligand Selection423
- IV. Applications424
- V. Conclusion427
- References428
- Chapter 11. Expanded-Bed Adsorption Process for Protein Capture431
- I. Introduction431
- II. Principles of Expanded-Bed Operation433
- III. Experimental Strategy435
- IV. Instrumentation437
- V. Matrices438
- VI. Applications438
- VII. Discussion and Conclusions449
- References450
- Chapter 12. Adsorptive Membranes for Bioseparations453
- I. Introduction454
- II. Comparison of Membrane Chromatography to Traditional Chromatography458
- III. Scale-Up of Chromatography Membranes460
- IV. Applications of MA to Preparative Bioseparations464
- V. Conclusions470
- References471
- Chapter 13. Simulated Moving-Bed Chromatography for Biomolecules475
- I. Introduction475
- II. Basic Principle476
- III. Operating Conditions482
- IV. Main Applications and Developments490
- V. Practical Application: Separation of Sugars499
- VI. Conclusion506
- References508
- Chapter 14. Large-Scale Chromatographic Purification of Oligonucleotides511
- I. Introduction512
- II. General Purification Strategies for Oligonucleotides516
- III. Large-Scale Purification of Therapeutic Oligonucleotides519
- IV. Purification of Related Molecules„DNA Fragments, Plasmids, Ribozymes, and RNA529
- V. Economics of Oligonucleotide Purification530
- VI. Summary531
- References531
- Chapter 15. Separation of Antibodies by Liquid Chromatography535
- I. Introduction536
- II. Antibodies: An Overview538
- III. Biological Starting Material546
- IV. Prepurification551
- V. Purification of Antibodies by Liquid Chromatography556
- VI. Regulatory Considerations612
- VII. General Conclusion on Antibody Separation Technologies620
- References621
- Chapter 16. Processing Plants and Equipment633
- I. Introduction633
- II. Industries Using Bioseparations634
- III. Process-Scale Bioseparations636
- IV. Process-Scale Considerations653
- V. Summary656
- References657
- Chapter 17. Engineering Process Control of Bioseparation Processes659
- I. Need for Process Control in Bioseparations660
- II. Brief Overview of Current Control Methods661
- III. Application Examples662
- IV. Opportunities for Continuing Development664
- References664
- Chapter 18. Economics of Bioseparation Processes667
- I. Introduction667
- II. Drugs Market and Sales670
- III. Applications of Models and Flow Sheets in Bioseparation Economics673
- References684
- Chapter 19. Future Developments687
- I. Introduction687
- II. The Partnership of Proteins and Nucleic Acids690
- III. Biotech Drugs690
- IV. Assuring Production and Purity694
- V. Genomics698
- VI. Lab on Chip699
- VII. Recovery of Biological Products700
- References710
- INDEX713
It is generally recognized that the commercial success of biotechnology products is highly dependent on the successful development and application of high-powered separation and purification methods. In this practical and authoritative handbook, the separation of proteins, nucleic acids, and oligonucleotides from biological matrices is covered from analytical to process scales. Also included in a chapter on the separation of monoclonal antibodies, which have found numerous uses as therapeutic and diagnostic agents. Analytical techniques include an interesting montage of chromatographic methods, capillary electrophoresis, isoelectric focusing, and mass spectrometry. Among separation and purification methods, liquid-liquid distribution, displacement chromatography, expanded bed adsorption, membrane chromatography, and simulated moving bed chromatography are covered at length. Regulatory and economic considerations are addressed, as are plant and process equipment and engineering process control. A chapter on future developments highlights the application of DNA chip arrays as well as evolving methodologies for a large number of drugs that are under development for treatment of cancer, AIDS, rheumatoid arthritis, and Alzheimer's disease. Handbook of Bioseparations serves as an essential reference and guidebook for separation scientists working in the pharmaceutical and biotechnology industries, academia, and government laboratories.
Key Features
* Covers bioseparations of proteins, nucleic acids, and monoclonal antibodies
* Encompasses both analytical and process-scale methods
* Elucidates the importance of engineering process control
* Details selection of plant and process equipment
* Addresses economic considerations
* Discusses future developments
Key Features
* Covers bioseparations of proteins, nucleic acids, and monoclonal antibodies
* Encompasses both analytical and process-scale methods
* Elucidates the importance of engineering process control
* Details selection of plant and process equipment
* Addresses economic considerations
* Discusses future developments