Preface xiv
1 Biopharmaceuticals, an Introductory Overview 1
1.1 Introduction to Pharmaceutical Products 1
1.2 Genetic Engineering and the Advent of Biopharmaceuticals 7
1.3 Biopharmaceuticals: Current Status and Future Prospects 12
Sources of Additional Information 20
Some Journal Articles 21
2 Biopharmaceuticals: Discovery, Development, and Regulation 23
2.1 Introduction 23
2.2 Discovery of Biopharmaceuticals 24
2.3 The Impact of "Omics" and Related Technologies Upon Drug Discovery 26
2.4 Lead Discovery 34
2.5 Pharmacogenetics 34
2.6 The Influence of Artificial Intelligence Upon Drug Discovery 35
2.7 Lead Characterization, CMC Development, and QbD 36
2.8 Delivery of Biopharmaceuticals 39
2.9 Preclinical Studies 43
2.10 Pharmacokinetics and Pharmacodynamics 43
2.11 Toxicity Studies 50
2.12 Clinical Trials 53
2.13 The Role and Remit of Regulatory Authorities 57
2.14 The Food and Drug Administration (FDA) 58
2.15 European Pharmaceutical Law 61
2.16 Pharmacovigilance 65
2.17 World Harmonization of Drug Regulations and the ICH 66
2.18 The Naming of Biopharmaceuticals 67
2.19 Patenting 68
2.20 Biosimilar Regulation 69
Sources of Additional Information 73
Some Journal Articles 73
3 Good Manufacturing Practice 75
3.1 Introduction 75
3.2 Guides to Good Manufacturing Practice (GMP) 75
3.3 Pharmacopoeia 77
3.4 The Manufacturing Facility 78
3.5 Documentation 93
3.6 Validation 95
Further Reading 97
4 Protein Structure 99
4.1 Introduction 99
4.2 Primary Structure 99
4.3 Higher-Level Structure 106
4.4 Protein Folding and Stability 111
4.5 Protein Structure Prediction 113
4.6 Protein Post-translational Modification 115
Further Reading 123
5 Production of Therapeutic Proteins by Recombinant Means 125
5.1 Introduction 125
5.2 Nucleic Acids: Structure and Function 125
5.3 The Principles of Genetic Engineering 133
5.4 Cell Types in Which Recombinant Therapeutic Proteins Are Produced 147
5.5 Engineered Therapeutic Proteins 159
Further Reading 164
6 Therapeutic Protein Manufacture: Upstream Processing 165
6.1 Introduction 165
6.2 Cell Banking Systems 166
6.3 Bioreactors 168
6.4 Microbial Cell Fermentation 174
6.5 Mammalian Cell Culture 176
Further Reading 180
7 Therapeutic Protein Manufacture: Downstream Processing 181
7.1 Initial Product Recovery 181
7.2 Initial Product Concentration 187
7.3 Chromatographic Purification 190
7.4 Viral Clearance 212
7.5 Some Influences Which can Alter the Biological Activity of Proteins 213
7.6 Final Product Formulation 217
7.7 Sterilization and Final Product Fill 219
7.8 Trends in Downstream Processing 224
Further Reading 225
8 Product Analysis 227
8.1 Introduction and Regulatory Context 227
8.2 Potency Determination 229
8.3 Protein Content 232
8.4 Purity Determination and API Characterization 233
8.5 Key Analytical Methodologies 235
8.6 Product Purity Determination 238
8.7 API Characterization 240
8.8 Non- protein Impurity Characterization: DNA 247
8.9 Contaminant Testing 248
8.10 Additional Tests 255
Further Reading 255
9 Antibodies 257
9.1 Antibody Structure and Function 257
9.2 Polyclonal Antibody Preparations Used Therapeutically 262
9.3 Hybridoma Technology and First- generation Monoclonal Antibodies 265
9.4 Chimeric and Humanized Antibodies 268
9.5 Fully Human mAbs 271
9.6 Routine Manufacture of mAbs 274
9.7 mAb Therapeutic Applications in Overview 275
9.8 mAb Therapeutics Treating Inflammation 275
9.9 mAbs Therapeutic Treating Cancer 277
9.10 Some Additional Therapeutic mAbs 283
9.11 Additional Engineered Antibody Formats 286
9.12 The mAb Market and the Advent of mAb Biosimilars 302
Further Reading 305
10 Vaccines 307
10.1 Introduction 307
10.2 Traditional Vaccine Preparations 309
10.3 The Impact of Recombinant DNA Technology on Vaccines 313
10.4 Adjuvant Technology 332
Further Reading 335
11 The Cytokines - The Interferon Family 337
11.1 Introduction to the Cytokines 337
11.2 The Interferons 344
11.3 Interferon Biotechnology 356
Further Reading 363
12 The Cytokines - Interleukins and Tumor Necrosis Factor 365
12.1 Introduction to the Interleukins 365
12.2 Interleukin- 2 367
12.3 IL- 2 Biotechnology 370
12.4 Interleukin 11 373
12.5 Inhibition of Interleukin Activity 374
12.6 Tumor Necrosis Factor 379
Further Reading 389
13 Growth Factors 391
13.1 Introduction 391
13.2 Hematopoietic Growth Factors 391
13.3 Granulocyte Colony- Stimulating Factor (G- CSF) 394
13.4 Macrophage colony- stimulating factor (M- CSF) 395
13.5 Granulocyte Macrophage Colony- Stimulating Factor (GM- CSF) 395
13.6 Clinical Application of CSFs 396
13.7 Erythropoietin 400
13.8 Therapeutic Applications of EPO 402
13.9 Thrombopoietin 404
13.10 Insulin- Like Growth Factors 407
13.11 Neurotrophic Factors 408
13.12 Fibroblast Growth Factors 410
13.13 Platelet- Derived Growth Factor 411
13.14 Bone Morphogenetic Proteins 412
13.15 Inhibition of Growth Factor Activity 413
Further Reading 419
14 Hormones 421
14.1 Introduction 421
14.2 Insulin 421
14.3 The Insulin Molecule 423
14.4 The Insulin Receptor and Signal Transduction 424
14.5 Insulin Manufacture by Traditional Means 426
14.6 Production of Human Insulin by Recombinant DNA Technology 427
14.7 Formulation of Insulin Products 428
14.8 Engineered Insulins 430
14.9 Glucagon- Like Peptide- 1 (GLP- 1)- Related Products 434
14.10 GLP- 1 Receptor Agonists 436
14.11 Human Growth Hormone 440
14.12 The Gonadotrophins 444
14.13 Medical Applications of Gonadotrophins 451
14.14 Additional Recombinant Hormones Now Approved 455
Further Reading 458
15 Recombinant Blood Products and Therapeutic Enzymes 459
15.1 Introduction 459
15.2 Hemostasis 459
15.3 The Coagulation Pathway 459
15.4 Clotting Disorders 463
15.5 Factor VIII and Hemophilia A 464
15.6 Additional Recombinant Blood Factors 470
15.7 Anticoagulants 473
15.8 Thrombolytic Agents 475
15.9 Enzymes of Therapeutic Value 480
Further Reading 492
16 Nucleic- Acid- Based Products 493
16.1 Introduction 493
16.2 Classical Gene Therapy 494
16.3 Basic Approach to Gene Delivery 495
16.4 Vectors Used in Gene Therapy 499
16.5 Manufacture of Viral Vectors 510
16.6 CAR- T- Based Gene Therapy 518
16.7 Non- viral Vectors 522
16.8 The Impact of CRISPR Technology on Gene Therapy 524
16.9 Antisense Technology, Including siRNA 527
Further Reading 533
Index 535
