Pharmaceutical Biotechnology - Concepts And Applications

Pharmaceutical Biotechnology offers students taking Pharmacy and related Medical and Pharmaceutical courses a comprehensive introduction to the fast-moving area of biopharmaceuticals. With a particular focus on the subject taken from a pharmaceutical perspective, initial chapters offer a broad introduction to protein science and recombinant DNA technology- key areas that underpin the whole subject. Subsequent chapters focus upon the development, production and analysis of these substances. Finally the book moves on to explore the science, biotechnology and medical applications of specific biotech products categories. These include not only protein-based substances but also nucleic acid and cell-based products.
introduces essential principles underlining modern biotechnology- recombinant DNA technology and protein science
an invaluable introduction to this fast-moving subject aimed specifically at pharmacy and medical students
includes specific ‘product category chapters’ focusing on the pharmaceutical, medical and therapeutic properties of numerous biopharmaceutical products.
entire chapter devoted to the principles of genetic engineering and how these drugs are developed.
includes numerous relevant case studies to enhance student understanding
no prior knowledge of protein structure is assumed

1 Pharmaceuticals, Biologics and Biopharmaceuticals
1.1 Introduction to pharmaceutical products
1.2 Biopharmaceuticals and pharmaceutical biotechnology
1.3 History of the pharmaceutical industry
1.4 The age of biopharmaceuticals
1.5 Biopharmaceuticals: current status and future prospects
Further reading 2 Protein Structure
2.1 Introduction
2.2 Overview of protein structure
2.2.1 Primary structure
2.2.2 The peptide bond
2.2.3 Amino acid sequence determination
2.2.4 Polypeptide synthesis
2.3 Higher level structure
2.3.1 Secondary structure
2.3.2 Tertiary structure
2.3.3 Higher structure determination
2.4 Protein stability and folding
2.4.1 Structural prediction
2.5 Protein post-translational modification
2.5.1 Glycosylation
2.5.2 Carboxylation and hydroxylation
2.5.3 Sulfation and amidation
Further reading 3 Gene Manipulation and Recombinant DNA Technology
3.1 Introduction
3.2 Nucleic acids: function and structure
3.2.1 Genome and gene organization
3.2.2 Nucleic acid purification
3.2.3 Nucleic acid sequencing
3.3 Recombinant production of therapeutic proteins
3.4 Classical gene cloning and identification
3.4.1 cDNA cloning
3.4.2 Cloning via polymerase chain reaction
3.4.3 Expression vectors
3.4.4 Protein engineering
Further reading 4 The Drug Development Process
4.1 Introduction
4.2 Discovery of biopharmaceuticals
4.3 The impact of genomics and related technologies upon drug discovery
4.4 Gene chips
4.5 Proteomics
4.6 Structural genomics
4.7 Pharmacogenetics
4.8 Initial product characterization
4.9 Patenting
4.9.1 What is a patent and what is patentable?
4.9.2 Patenting in biotechnology
4.10 Delivery of biopharmaceuticals
4.10.1 Oral delivery systems
4.10.2 Pulmonary delivery
4.10.3 Nasal, transmucosal and transdermal delivery systems
4.11 Preclinical studies
4.12 Pharmacokinetics and pharmacodynamics
4.12.1 Protein pharmacokinetics
4.12.2 Tailoring of pharmacokinetic profile
4.12.3 Protein mode of action and pharmacodynamics
4.13 Toxicity studies
4.13.1 Reproductive toxicity and teratogenicity
4.13.2 Mutagenicity, carcinogenicity and other tests
4.13.3 Clinical trials
4.13.4 Clinical trial design
4.13.5 Trial size design and study population
4.14 The role and remit of regulatory authorities
4.14.1 The Food and Drug Administration
4.14.2 The investigational new drug application
4.14.3 The new drug application
4.14.4 European regulations
4.14.5 National regulatory authorities
4.14.6 The European Medicines Agency and the new EU drug approval systems
4.14.7 The centralized procedure
4.14.8 Mutual recognition
4.14.9 Drug registration in Japan
4.14.10 World harmonization of drug approvals
4.14 Conclusion
Further reading 5 Sources and Upstream Processing
5.1 Introduction
5.2 Sources of biopharmaceuticals
5.2.1 Escherichia coli as a source of recombinant, therapeutic proteins
5.2.2 Expression of recombinant proteins in animal cell culture systems
5.2.3 Additional production systems
5.2.3.1 Yeast
5.2.3.2 Fungal production systems
5.2.3.3 Transgenic animals
5.2.3.4 Transgenic plants
5.2.3.5 Insect cell-based systems
5.3 Upstream processing
5.3.1 Cell banking systems
5.3.2 Microbial cell fermentation
5.3.3 Mammalian cell culture systems
Further reading 6 Downstream Processing
6.1 Introduction
6.2 Initial product recovery
6.3 Cell disruption
6.4 Removal of nucleic acid
6.5 Initial product concentration
6.5.1 Ultrafiltration
6.5.2 Diafiltration
6.6 Chromatographic purification
6.6.1 Size-exclusion chromatography (gel filtration)
6.6.2 Ion-exchange chromatography
6.6.3 Hydrophobic interaction chromatography
6.6.4 Affinity chromatography
6.6.5 Immunoaffinity purifications
6.6.6 Protein A chromatography
6.6.7 Lectin affinity chromatography
6.6.8 Dye affinity chromatography
6.6.9 Metal chelate affinity chromatography
6.6.10 Chromatography on hydroxyapatite
6.6.11 Chromatofocusing
6.7 High-performance liquid chromatography of proteins
6.8 Purification of recombinant proteins
6.9 Final product formulation
6.9.1 Some influences that can alter the biological activity of proteins
6.9.1.1 Proteolytic degradation and alteration of sugar side chains.
6.9.1.2 Protein deamidation
6.9.1.3 Oxidation and disulfide exchange
6.9.2 Stabilizing excipients used in final product formulations
6.9.3 Final product fill
6.9.4 Freeze-drying
6.9.5 Labelling and packing
Further reading 7 Product Analysis
7.1 Introduction
7.2 Protein-based contaminants
7.3 Removal of altered forms of the protein of interest from the product stream
7.3.1 Product potency
7.3.2 Determination of protein concentration
7.4 Detection of protein-based product impurities
7.4.1 Capillary electrophoresis
7.4.2 High-pressure liquid chromatography
7.4.3 Mass spectrometry
7.5 Immunological approaches to detection of contaminants
7.5.1 Amino acid analysis
7.5.2 Peptide mapping
7.5.3 N-terminal sequencing
7.5.4 Analysis of secondary and tertiary structure
7.6 Endotoxin and other pyrogenic contaminants
7.6.1 Endotoxin, the molecule
7.6.2 Pyrogen detection
7.6.3 DNA
7.6.4 Microbial and viral contaminants
7.6.5 Viral assays
7.6.6 Miscellaneous contaminants
7.6.7 Validation studies
Further reading 8 The Cytokines: The Interferon Family
8.1 Cytokines
8.1.1 Cytokine receptors
8.1.2 Cytokines as biopharmaceuticals
8.2 The interferons
8.2.1 The biochemistry of interferon-a
8.2.2 Interferon-b
8.2.3 Interferon-g
8.2.4 Interferon signal transduction
8.2.5 The interferon receptors
8.2.6 The JAK–STAT pathway
8.2.7 The interferon JAK–STAT pathway
8.2.8 The biological effects of interferons
8.2.9 The eIF-2a protein kinase system
8.3 Interferon biotechnology
8.3.1 Production and medical uses of interferon-a
8.3.2 Medical uses of interferon-b
8.3.3 Medical applications of interferon-g
8.3.4 Interferon toxicity
8.3.5 Additional interferons
8.4 Conclusion
Further reading 9 Cytokines: Interleukins and Tumour Necrosis Factor
9.1 Introduction
9.2 Interleukin-2
9.2.1 Interleukin-2 production
9.2.2 Interleukin-2 and cancer treatment
9.2.3 Interleukin-2 and infectious diseases
9.2.4 Safety issues
9.2.5 Inhibition of interleukin-2 activity
9.3 Interleukin-1
9.3.1 The biological activities of interleukin-1
9.3.2 Interleukin-1 biotechnology
9.4 Interleukin-11
9.5 Tumour necrosis factors
9.5.1 Tumour necrosis factor biochemistry
9.5.2 Biological activities of tumour necrosis factor-a
9.5.3 Immunity and inflammation
9.5.4 Tumour necrosis factor receptors
9.5.5 Tumour necrosis factor: therapeutic aspects
Further reading 10 Growth Factors
10.1 Introduction
10.2 Haematopoietic growth factors
10.2.1 The interleukins as haemopoietic growth factors
10.2.2 Granulocyte colony-stimulating factor
10.2.3 Macrophage colony-stimulating factor
10.2.4 Granulocyte macrophage colony-stimulating factor
10.2.5 Clinical application of colony-stimulating factors
10.2.6 Erythropoietin
10.2.6.1 Therapeutic applications of erythropoietin
10.2.6.2 Chronic disease and cancer chemotherapy
10.2.7 Thrombopoietin
10.3 Growth factors and wound healing
10.3.1 Insulin-like growth factors
10.3.2 Insulin-like growth factor biological effects
10.3.3 Epidermal growth factor
10.3.4 Platelet-derived growth factor
10.3.5 Fibroblast growth factors
10.3.6 Transforming growth factors
10.3.7 Neurotrophic factors
Further reading 11 Therapeutic Hormones
11.1 Introduction
11.2 Insulin
11.2.1 Diabetes mellitus
11.2.2 The insulin molecule
11.2.3 The insulin receptor and signal transduction
11.2.4 Insulin production
11.2.5 Production of human insulin by recombinant DNA technology
11.2.6 Formulation of insulin products
11.2.7 Engineered insulins
11.2.8 Additional means of insulin administration
11.3 Glucagon
11.4 Human growth hormone
11.4.1 The growth hormone receptor
11.4.2 Biological effects of growth hormone
11.4.3 Therapeutic uses of growth hormone
11.5 The gonadotrophins
11.5.1 Follicle-stimulating hormone, luteinizing hormone and human chorionic gonadotrophin
11.5.2 Pregnant mare serum gonadotrophin
11.5.3 The inhibins and activins
11.6 Medical and veterinary applications of gonadotrophins
11.6.1 Sources and medical uses of follicle-stimulating hormone, luteinizing hormone and human chorionic gonadotrophin
11.6.2 Recombinant gonadotrophins
11.6.3 Veterinary uses of gonadotrophins
11.7 Additional recombinant hormones now approved
11.8 Conclusion
Further reading 12 Recombinant Blood Products and Therapeutic Enzymes
12.1 Introduction
12.2 Haemostasis
12.2.1 The coagulation pathway
12.2.2 Terminal steps of coagulation pathway
12.2.3 Clotting disorders
12.2.4 Factor VIII and haemophilia
12.2.5 Production of factor VIII
12.2.6 Factors IX, IIVa and XIII
12.3 Anticoagulants
12.3.1 Hirudin
12.3.2 Antithrombin
12.4 Thrombolytic agents
12.4.1 Tissue plasminogen activator
12.4.2 First-generation tissue plasminogen activator
12.4.3 Engineered tissue plasminogen activator
12.4.4 Streptokinase
12.4.5 Urokinase
12.4.6 Staphylokinase
12.4.7 a1-Antitrypsin
12.4.8 Albumin
12.5 Enzymes of therapeutic value
12.5.1 Asparaginase
12.5.2 DNase
12.5.3 Glucocerebrosidase
12.5.4 a-Galactosidase, urate oxidase and laronidase
12.5.5 Superoxide dismutase
12.5.6 Debriding agents
12.5.7 Digestive aids
Further reading 13 Antibodies, Vaccines and Adjuvants
13.1 Introduction
13.2 Traditional polyclonal antibody preparations
13.3 Monoclonal antibodies
13.3.1 Antibody screening: phage display technology
13.3.2 Therapeutic application of monoclonal antibodies
13.3.3 Tumour immunology
13.3.3.1 Antibody-based strategies for tumour detection/destruction
13.3.3.2 Drug-based tumour immunotherapy
13.3.3.3 First-generation anti-tumour antibodies: clinical disappointment
13.3.4 Tumour-associated antigens
13.3.5 Antigenicity of murine monoclonals
13.3.6 Chimaeric and humanized antibodies
13.3.7 Antibody fragments
13.3.8 Additional therapeutic applications of monoclonal antibodies
13.4 Vaccine technology
13.4.1 Traditional vaccine preparations
13.4.1.1 Attenuated, dead or inactivated bacteria
13.4.1.2 Attenuated and inactivated viral vaccines
13.4.1.3 Toxoids and antigen-based vaccines
13.4.2 The impact of genetic engineering on vaccine technology
13.4.3 Peptide vaccines
13.4.4 Vaccine vectors
13.4.5 Development of an AIDS vaccine
13.4.6 Difficulties associated with vaccine development
13.4.7 AIDS vaccines in clinical trials
13.4.8 Cancer vaccines
13.4.9 Recombinant veterinary vaccines
13.5 Adjuvant technology
13.5.1 Adjuvant mode of action
13.5.2 Mineral-based adjuvants
13.5.3 Oil-based emulsion adjuvants
13.5.4 Bacteria/bacterial products as adjuvants
13.5.5 Additional adjuvants
Further reading 14 Nucleic-Acid- and Cell-Based Therapeutics
14.1 Introduction
14.2 Gene therapy
14.2.1 Basic approach to gene therapy
14.2.2 Some additional questions
14.3 Vectors used in gene therapy
14.3.1 Retroviral vectors
14.3.2 Adenoviral and additional viral-based vectors
14.3.3 Manufacture of viral vectors
14.3.4 Non-viral vectors
14.3.5 Manufacture of plasmid DNA
14.4 Gene therapy and genetic disease
14.5 Gene therapy and cancer
14.6 Gene therapy and AIDS
14.6.1 Gene-based vaccines
14.6.2 Gene therapy: some additional considerations
14.7 Antisense technology
14.7.1 Antisense oligonucleotides and their mode of action
14.7.2 Uses, advantages and disadvantages of ‘oligos’
14.8 Oligonucleotide pharmacokinetics and delivery
14.8.1 Manufacture of oligos
14.8.2 Additional antigene agents: RNA interference and ribozymes
14.9 Aptamers
14.10 Cell- and tissue-based therapies
14.10.1 Stem cells
14.10.2 Adult stem cells.
14.11 Conclusion
Further reading
Index