Pharmaceutical Biotechnology: A Focus on Industrial Application

Cover\nHalf Title\nTitle Page\nCopyright Page\nTable of Contents\nPreface\nEditors\nContributors\nChapter 1 Fundamentals of Biotechnology\n	1.1 Introduction\n	1.2 Biological Molecules\n		1.2.1 Introduction\n		1.2.2 Proteins\n		1.2.3 Nucleic Acids\n		1.2.4 Virus\n	1.3 Basic Technologies of the Biotechnological Processes\n		1.3.1 Monoclonal Antibody Technology\n		1.3.2 Bioprocessing Technology\n		1.3.3 Cell Culture Technology\n		1.3.4 Tissue Engineering Technology\n		1.3.5 Biosensors Technology\n		1.3.6 Genetic Engineering Technology\n		1.3.7 Protein Engineering Technology\n		1.3.8 Antisense RNA Technology\n		1.3.9 Chip DNA Technology\n		1.3.10 Biocomputing Technology\n	1.4 Biotechnology and Applications\n		1.4.1 Medical\n			1.4.1.1 Diagnostics\n			1.4.1.2 Therapeutics\n		1.4.2 Environment\n		1.4.3 Livestock Raise and Agriculture\n	1.5 Matters of Business\n		1.5.1 Structuring of a Biotechnology Enterprise\n		1.5.2 Biopharmaceutical Industry\n	1.6 Social Aspects of Biotechnology\n		1.6.1 Use of Bioproducts\n		1.6.2 Genetic Privacy and Laboratory Prognostic Testing\n		1.6.3 Stem Cells and Cloning\n		1.6.4 Use of Test Subjects\n		1.6.5 Agriculture\n	1.7 Final Considerations\n	References\nChapter 2 Thermodynamics Applied to Biomolecules\n	2.1 Introduction\n	2.2 Thermodynamic Concepts\n	2.3 Mathematical Modelling\n		2.3.1 Reaction Thermodynamics\n			2.3.1.1 Activation Energy and Standard Enthalpy Variation of the Unfolding Equilibrium\n			2.3.1.2 Enthalpy, Entropy and Gibbs Free Energy of the Reaction\n		2.3.2 Thermodynamics of Biomolecules Thermo-inactivation\n			2.3.2.1 Enthalpy, Entropy and Gibbs Free Energy of Biocatalyst Thermo-Inactivation\n			2.3.2.2 Complementary Kinetic Parameters\n	2.4 Experimental Procedures and Case Studies\n		2.4.1 Thermodynamics of Enzyme-Catalysed Reactions\n			2.4.1.1 Experimental Procedure\n			2.4.1.2 Practical Example\n		2.4.2 Thermodynamics of Biocatalyst Thermal Denaturation\n			2.4.2.1 Experimental Procedure\n			2.4.2.2 Practical Example\n	2.5 Physical and Chemical Factors Influencing Thermodynamic Parameters\n		2.5.1 Temperature\n		2.5.2 pH\n		2.5.3 Substrate Concentration\n		2.5.4 Techniques of Biomolecule Improvement\n	2.6 Final Considerations\n	References\nChapter 3 Expression Systems for the Production of Therapeutic Recombinant Proteins\n	3.1 Introduction\n	3.2 Differences between Synthetic Drugs and Biopharmaceuticals Based on Recombinant Proteins\n	3.3 Main Systems for Expression of Therapeutic Recombinant Proteins\n		3.3.1 Bacteria\n		3.3.2 Yeasts\n		3.3.3 Insect Cells\n		3.3.4 Plant Cells\n		3.3.5 Mammalian Cells\n	3.4 Development of Bioprocesses for the Production of Therapeutic Recombinant Proteins\n	3.5 Final Considerations\n	References\nChapter 4 Molecular Biology: Tools in Industrial Pharmaceutical Biotechnology\n	4.1 Introduction\n	4.2 Therapeutic Targets\n	4.3 Cellular Communication\n	4.4 Expression Gene Regulation\n	4.5 Transcriptional System in Prokaryotes\n		4.5.1 Promoter Structure and Operon\n		4.5.2 Transcription Termination\n	4.6 Transcription Regulation in Eukaryotes\n	4.7 RNA Processing\n	4.8 Final Considerations\n	References\nChapter 5 Molecular Biology Tools: Techniques and Enzymes\n	5.1 Introduction\n	5.2 Isolating a Gene Sequence-DNA Polymerase\n	5.3 Manipulating the Gene Obtained by PCR\n	5.4 Restriction Enzymes and Expression Vectors\n	5.5 Final Considerations\n	References\nChapter 6 Bioinformatics Applied to the Development of Biomolecules of Pharmaceutical Interest\n	6.1 Introduction\n	6.2 Databases\n		6.2.1 Databases of Small Molecules\n		6.2.2 Natural Products Databases\n		6.2.3 Nucleic Acid Databases\n		6.2.4 Peptide Databases\n		6.2.5 Protein Databases\n		6.2.6 Other Relevant Biological Databases\n	6.3 Structural Protein Analysis\n		6.3.1 Visualization Programmes and Analysis of Three-Dimensional Structures\n		6.3.2 Protein Modelling. Why Model?\n			6.3.2.1 Basis of Methods\n			6.3.3.1 MD\n			6.3.3.2 NMA: Normal Modes Analysis\n			6.3.3.3 Comparison: MD vs NMA\n			6.3.3.4 Use\n			6.3.3.5 Interaction Models: Protein vs Ligand\n			6.3.3.6 Docking: Protein vs Small Molecule\n			6.3.3.7 Docking: Protein vs Protein\n	6.4 Examples\n		6.4.1 Bioinformatics in Vaccine Discovery\n		6.4.2 Use of Peptides in New Drug Development\n		6.4.3 Human Immunodeficiency Virus Reverse Transcriptase Inhibitors\n		6.4.4 Applied Bioinformatics Synthetic Biology: Production and Discovery of New Biopharmaceuticals\n	6.5 Final Considerations\n	References\nChapter 7 Bioprocesses: Microorganisms and Culture Media\n	7.1 Introduction\n	7.2 General Characteristics of Microorganisms\n		7.2.1 Cell Concentration Measurement\n		7.2.2 Standard Dry Mass Curve\n			7.2.2.1 Results\n		7.2.3 Duplication and Generation Time\n	7.3 Cultivation Media of Industrial Interest\n		7.3.1 Trace Elements and Minerals\n		7.3.2 Growth Factors\n		7.3.3 Synthetic or Chemically Defined Media\n		7.3.4 Complex Media\n	7.4 Antifoam Agents\n	7.5 pH Control in the Bioprocess\n	7.6 Precursors\n	7.7 Primary and Secondary Metabolites\n		7.7.1 Overproduction of Secondary Metabolites\n	7.8 Final Considerations\n	References\nChapter 8 Sterilization in Pharmaceutical Biotechnology\n	8.1 Introduction\n	8.2 Terms and Definitions\n	8.3 Types of Sterilization\n		8.3.1 Physical Sterilization Methods\n			8.3.1.1 Heat\n			8.3.1.2 Ionizing Radiation\n			8.3.1.3 Filtration\n		8.3.2 Chemical Sterilization Methods\n	8.4 Sterilization in Upstream Operations\n		8.4.1 Culture Media and Its Sterilization\n		8.4.2 Batch Heat Sterilization\n		8.4.3 Heat Sterilization in a Continuous System\n		8.4.4 Sterilization Kinetics of Microorganisms\n		8.4.5 Air Sterilization\n	8.5 Sterilization in Downstream Operations\n		8.5.1 Terminal Sterilization and Aseptic Processing\n		8.5.2 Sterilization of Biopharmaceuticals in Downstream Processes\n		8.5.3 Sterilization of Biopharmaceuticals\n	8.6 Validation of Sterilization Processes\n	8.7 Final Considerations\n	References\nChapter 9 Kinetics of Cell Cultivation\n	9.1 Introduction\n	9.2 Typical Batch Cultivation Phases\n	9.3 With What Does μ Vary? (Model for Cell Growth)\n	9.4 What Does S Vary with? (Model for Substrate Consumption)\n	9.5 How to Calculate the Productivity of a Process?\n	9.6 How Does the Product Concentration Vary (Model for Production)?\n	9.7 Final Considerations\n	References\nChapter 10 Bioreactors: Modes of Operation\n	10.1 Introduction\n	10.2 General Characteristics\n		10.2.1 Batch Mode\n		10.2.2 Fed-Batch Mode\n		10.2.3 Continuous Mode\n	10.3 Mass Balances for Different Modes of Operation\n	10.4 Batch Operation\n		10.4.1 Mass Balance for the Cells in the Batch Mode\n		10.4.2 Mass Balance for the Limiting Substrate in the Batch\n		10.4.3 Mass Balance for the Product in Batch Mode\n	10.5 Fed-Batch Operation\n		10.5.1 Mass Balance for Cells in Fed-Batch Mode\n		10.5.2 Mass Balance for the Limiting Substrate in Fed-Batch Mode\n		10.5.3 Mass Balance for the Product in Fed-Batch Mode\n	10.6 Continuous Mode Operation\n		10.6.1 Mass Balance for Cells in Continuous Mode without Cell Recycling\n		10.6.2 Mass Balance for a Limiting Substrate in Continuous Mode without Cell Recycling\n		10.6.3 Mass Balance for the Product in Continuous Mode without Cell Recycling\n		10.6.4 Mass Balance for Cells in Continuous Mode with External Cell Recycling\n		10.6.5 Mass Balance for the Limiting Substrate in Continuous Mode with External Cell Recycling\n		10.6.6 Mass Balance for the Product in Continuous Mode with External Cell Recycling\n		10.6.7 How to Calculate the Volumetric Productivity of a Continuous Process?\n	10.7 Final Considerations\n	References\nChapter 11 Agitation and Aeration: Oxygen Transfer and Cell Respiration\n	11.1  Introduction\n	11.2  Gas Pressure and Oxygen Partial Pressure\n	11.3  Aeration Systems\n		11.3.1 Superficial Aeration\n		11.3.2 Submerged Aeration\n			11.3.2.1 Aeration by Sparging with Mechanical Agitation (Stirred Tank Reactor)\n			11.3.2.2 Aeration by Bubbling with No Mechanical Agitation\n	11.4  Mechanical Agitation Systems in Bioreactors\n	11.5  Oxygen Transfer and Respiration: From Gas Bubbles to Cells\n	11.6  Gas-Liquid Mass Transfer (Oxygen Supply): OTR\n	11.7  Oxygen Uptake in Cell Cultures (Oxygen Demand): OUR\n	11.8  Integrating Oxygen Supply and Demand\n	11.9  Dissolved Oxygen Concentration (C) Profile in a Culture with Constant Oxygen Transfer Conditions\n	11.10 Control of C\n	11.11 Measurement of k[sub(L)]a\n		11.11.1 Dynamic Method without Cells\n	11.12 Measurement of Q[sub(O2] X\n		11.12.1 Dynamic Method\n		11.12.2 Gas Balance\n		11.12.3 Liquid Phase Mass Balance\n	11.13 Criteria for Scaling Up the Bioprocess\n	11.14 Final Considerations\n	References\nChapter 12 Mammalian Cell Culture Technology\n	12.1 Introduction\n	12.2 Brief History\n	12.3 Main Product Categories\n		12.3.1 Vaccines\n		12.3.2 Monoclonal Antibodies\n		12.3.3 Glycoproteins\n		12.3.4 Cells and Tissues\n		12.3.5 Gene Therapy\n	12.4 Mammalian Cells\n		12.4.1 Basic Characteristics\n		12.4.2 Post-Translational Processes\n		12.4.3 Cell Types\n		12.4.4 Hybridoma Cells\n		12.4.5 Cell Culture Derivatives\n			12.4.5.1 Immunogenicity of Recombinant Proteins\n			12.4.5.2 Basic Structure of Antibodies or Immunoglobulins\n			12.4.5.3 Types of Monoclonal Antibodies\n			12.4.5.4 Phage Display Technology\n			12.4.5.5 Culture Media Used in Mammalian Cell Technology\n			12.4.5.6 Culture Medium Optimization\n		12.4.6 Basic Cell Culture Laboratory\n		12.4.7 Bioprocess and Mammalian Cells\n		12.4.8 Main Guidelines for Obtaining Cell Banks\n			12.4.8.1 Obtaining and Controlling the ‘Original’ Cell\n			12.4.8.2 Cell Bank Preparation\n			12.4.8.3 Qualification Tests of the Cell Bank\n			12.4.8.4 Karyotyping (Cytogenetic Analysis)\n			12.4.8.5 Analysis of Isoenzymes\n			12.4.8.6 DNA Fingerprinting\n			12.4.8.7 Genetic Stability of the Recombinant Mammalian Cell\n		12.4.9 Best Practices in Cell Culture\n		12.4.10 Development of a Cell Line\n			12.4.10.1 Transient Expression\n			12.4.10.2 Stable Expression\n			12.4.10.3 Construction of Expression Vector\n			12.4.10.4 Promoters\n			12.4.10.5 Enhancer\n			12.4.10.6 Elements that Stabilize and Increase the Translation of the Primary Transcript\n			12.4.10.7 Selection Markers\n			12.4.10.8 Selection of Clones\n			12.4.10.9 Transfection\n	12.5 Production Scale-Up\n	12.6 Bioreactors and Mammalian Cell Cultivation\n		12.6.1 Small Scale Culture\n		12.6.2 Scaling-Up Problems\n	12.7  Anchorage-Dependent Cell Systems\n		12.7.1 Roller Bottles\n		12.7.2 Stacked-Plate or Multitray Systems\n		12.7.3 Microcarriers\n		12.7.4 Bed Bioreactors\n	12.8  Systems for Cells in Suspension (or Cells Attached to Microcarriers)\n		12.8.1 Spinner Bottles\n		12.8.2 Shake Flasks\n		12.8.3 Culture Bags\n		12.8.4 Single-Use Bioreactors (SUBs)\n	12.9  Bioreactors\n		12.9.1 Hollow Fibre Bioreactors Applied to the Cultivation of Mammalian Cells\n		12.9.2 Type of Bioprocesses in Cell Culture\n		12.9.3 Cell Metabolism\n		12.9.4 Glucose, Glutamine and Amino Acids as Sources of Energy and Carbon\n		12.9.5 Effects of Lactate and Ammonia\n		12.9.6 Role of Oxygen and CO[sub(2)] in Cellular Metabolism of Mammalian Cells\n	12.10 Monitoring and Control of Mammalian Cell Culture\n		12.10.1 Partial Pressure of O[sub(2)] (pO[sub(2)])\n		12.10.2 Partial Pressure of Carbon Dioxide\n		12.10.3 Metabolites and Products\n		12.10.4 Cell Concentration and Viability\n	12.11 Final Considerations\n	References\nChapter 13 Purification Process of Biomolecules\n	13.1 Introduction\n	13.2 Cell-Liquid Separation\n		13.2.1 Filtration\n		13.2.2 Centrifugation\n	13.3 Cell Disruption\n	13.4 Concentration of Biomolecules\n		13.4.1 Precipitation\n		13.4.2 Tangential Filtration\n		13.4.3 Liquid-Liquid Extraction\n	13.5 Chromatographic Processes\n		13.5.1 Molecular Exclusion\n		13.5.2 Ion Exchange\n		13.5.3 Hydrophobic Interaction\n		13.5.4 Affinity Chromatography\n		13.5.5 Scale Up\n		13.5.6 Expanded Bed Adsorption\n	13.6 Final Treatment\n	13.7 Purification of Monoclonal Antibodies\n	13.8 Endotoxin Removal\n	13.9 Yield and Purity\n	13.10 Identification of Biomolecules\n	13.11 Trends in Processes Applied to the Purification of Biomolecules\n	13.12 Final Considerations\n	References\nChapter 14 Lipopolysaccharides: Methods of Quantification and Removal from Biotechnological Products\n	14.1 Introduction\n	14.2 LPS Characteristics and Properties\n	14.3 LPS Functions and Mechanisms of Action\n	14.4 Main Techniques for LPS Quantification in Biotechnological Products\n		14.4.1 LPS Test in Rabbits\n		14.4.2 LPS Test – Gel Clot\n		14.4.3 LPS Test – Chromogenic and Turbidimetric Kinetics\n		14.4.4 New Methods for LPS Quantification\n	14.5 Techniques Used to Remove LPS from Biotechnological Products\n		14.5.1 Chromatographic Techniques\n			14.5.1.1 Affinity Chromatography\n			14.5.1.2 Ion-Exchange Chromatography\n			14.5.1.3 Hydrophobic Interaction Chromatography\n		14.5.2 Membrane-Based Filtration\n		14.5.3 Ultrafiltration\n		14.5.4 Microfiltration\n		14.5.5 Depth Filtration\n		14.5.6 Novel Methods for LPS Removal\n			14.5.6.1 Aqueous Two-Phase Systems\n	14.6 Final Considerations\n	References\nChapter 15 Enzymes: The Catalytic Proteins\n	15.1 Introduction\n	15.2 Enzyme Specificity\n	15.3 Enzyme Activity\n		15.3.1 Quantification of Enzyme Activity\n		15.3.2 Expression of Enzyme Activity\n		15.3.3 Factors Affecting Enzyme Activity\n			15.3.3.1 Physical-Chemical Factors\n			15.3.3.2 Chemical Factors\n			15.3.3.3 Physical Factors\n		15.3.4 Thermodynamics of Enzyme Catalysis\n	15.4 Final Considerations\n	References\nChapter 16 Enzymes as Drugs and Medicines\n	16.1 Introduction\n	16.2 Enzymes in Medicines\n		16.2.1 Enzymes Bioavailability\n			16.2.1.1 Transport across Endothelium\n		16.2.2 Therapeutic Enzyme Delivery\n		16.2.3 Acylation of Enzymes\n		16.2.4 Molecular Modelling of Enzymes\n		16.2.5 Important Aspects of Therapeutic Enzymes\n	16.3 Enzymes in Clinical Analysis and Cosmetics\n		16.3.1 Clinical Analysis\n		16.3.2 Cosmetics\n	16.4 Final Considerations\n	References\nChapter 17 Aspects of the Immobilization Technique\n	17.1 Introduction\n	17.2 Types of Immobilization\n		17.2.1 Entrapment\n			17.2.1.1 Cross-Linked Matrices\n			17.2.1.2 Encapsulation\n			17.2.1.3 Microencapsulation\n		17.2.2 Bonding Formation\n			17.2.2.1 Adsorption\n			17.2.2.2 Covalent Binding\n			17.2.2.3 Cross-Linking\n		17.2.3 Supports\n		17.2.4 Effects Caused by Immobilization\n			17.2.4.1 Steric and Conformational Effects\n			17.2.4.2 Diffusion and Mass Transport Effects\n			17.2.4.3 Effects of Microenvironment\n			17.2.4.4 Advantages and Disadvantages of the Immobilization Technique\n			17.2.4.5 Applications\n	17.3 Fundamentals of Enzyme Reactors\n		17.3.1 Types of Enzyme Reactors\n		17.3.2 Enzyme Reactor Kinetics\n		17.3.3 Operation of Enzyme Reactors\n	17.4 Final Considerations\n	References\nChapter 18 Biomolecules in Analytical Methods\n	18.1 Introduction\n	18.2 Enzymes as Diagnostic Tools\n		18.2.1 Diagnosis of Gastric Disorders\n		18.2.2 Neonatal Screening\n		18.2.3 Cancer Diagnosis\n		18.2.4 Enzyme Detection of Micronutrients, Microorganisms and Cholesterol\n	18.3 Test Strips\n	18.4 Biosensors\n		18.4.1 General Characteristics\n		18.4.2 Electrochemical Detectors\n			18.4.2.1 Amperometric Biosensors\n			18.4.2.2 Potentiometric Biosensors\n			18.4.2.3 Optical Detectors\n			18.4.2.4 Thermal Detectors\n			18.4.2.5 Piezoelectric Detectors\n			18.4.2.6 Immunodetection Biosensors\n	18.5 Novel Trends in Biosensing Technology\n		18.5.1 Microfluidic Integrated Biosensors: Towards Lab-on-a-Chip\n		18.5.2 Wearable Biosensors for Healthcare Monitoring\n	18.6 Final Considerations\n	References\nChapter 19 Nanotechnology and Biopharmaceuticals\n	19.1  Introduction\n	19.2  Nanobiotechnology for Biopharmaceuticals\n	19.3  Liposomes\n	19.4  Polymersomes\n	19.5  Polymeric Micelles\n	19.6  Polymeric Nanoparticles: Nanocapsules and Nanospheres\n	19.7  Nanoemulsions and Microemulsions\n	19.8  Solid Lipid Nanoparticles and Nanostructured Lipid Carriers\n	19.9  PEGylation\n	19.10 Characterization Techniques\n		19.10.1 Dynamic Light Scattering (DLS)\n		19.10.2 Nanoparticle Tracking Analysis (NTA)\n		19.10.3 Zeta Potential (ZP)\n		19.10.4 Microscopy\n	19.11 Nanotoxicity\n	19.12 Regulatory Aspects\n	19.13 Final Considerations\n	References\nChapter 20 Biosafety Applied in Pharmaceutical and Biotechnological Processes\n	20.1 Introduction\n	20.2 Definitions of Biosafety\n	20.3 Classification of Microorganisms by Risk Group\n	20.4 Biosafety Levels\n		20.4.1 Biosafety Level 1 (BSL-1)\n			20.4.1.1 Laboratory Practices for BSL-1\n			20.4.1.2 Safety Equipment for BSL-1\n			20.4.1.3 BSL-1 Laboratory Facility\n		20.4.2 Biosafety Level 2 (BSL-2)\n			20.4.2.1 Laboratory Practices for BSL-2\n			20.4.2.2 Safety Equipment for BSL-2\n			20.4.2.3 BSL-2 Laboratory Facility\n		20.4.3 Biosafety Level 3 (BSL-3)\n			20.4.3.1 Laboratory Practices for BSL-3\n			20.4.3.2 Safety Equipment for BSL-3\n			20.4.3.3 BSL-3 Laboratory Facility\n		20.4.4 Biosafety Level 4 (BSL-4)\n			20.4.4.1 Laboratory Practices for BSL-4\n			20.4.4.2 Safety Equipment for BSL-4\n			20.4.4.3 BSL-4 Laboratory Facility\n	20.5 Laboratory Accidents\n		20.5.1 Historical Accidents\n		20.5.2 Accident Prevention\n			20.5.2.1 Proper Laboratory Supervision\n			20.5.2.2 Training and Awareness\n			20.5.2.3 Continued Education\n			20.5.2.4 Proper Laboratory Techniques\n			20.5.2.5 Proper Laboratory Equipment\n			20.5.2.6 Laboratory Organization\n			20.5.2.7 Vaccination\n	20.6 Activity Planning\n	20.7 Safety Equipment\n	20.8 Final Considerations\n	References\nChapter 21 Pharmaceutical Quality System for Biotechnology Products\n	21.1 Introduction\n	21.2 Pharmaceutical Quality System (PQS)\n	21.3 Quality-by-Design in Biotechnology Product and Process Development\n		21.3.1 Historical Context\n		21.3.2 QbD-based Development\n			21.3.2.1 Product Design Space\n			21.3.2.2 Process Design Space\n			21.3.2.3 Control Strategy\n			21.3.2.4 Lifecycle Management Plan\n	21.4 Practical Notes on Process Development\n		21.4.1 Expression Systems\n			21.4.1.1 Bacterial Systems\n			21.4.1.2 Filamentous Fungi and Yeast Systems\n			21.4.1.3 Mammalian Cell Systems\n		21.4.2 Where Does Biopharmaceutical Scale-Up Start?\n	21.5 Good Manufacturing Practices (GMP)\n		21.5.1 Validation – Production Process\n		21.5.2 Process Validation in Bioreactors (Upstream)\n		21.5.3 Validation of Extraction and Purification Processes (Downstream)\n		21.5.4 Validation – Viral Safety\n			21.5.4.1 Physical Methods\n			21.5.4.2 Chemical Methods\n		21.5.5 Validation – Analytical Methods\n			21.5.5.1 Reference Standard\n			21.5.5.2 Method Development\n		21.5.6 Validation – Biological Assays\n		21.5.7 Change Control\n	21.6 Analytical Methods for Laboratory Quality Control\n		21.6.1 Physicochemical Tests\n			21.6.1.1 High-Performance Liquid Chromatography (HPLC)\n			21.6.1.2 Gel Electrophoresis\n			21.6.1.3 NMR – Nuclear Magnetic Resonance\n			21.6.1.4 Capillary Electrophoresis\n			21.6.1.5 Spectrophotometric Methods\n		21.6.2 Others (pH, Total Solids, Preservatives, Isotonicity)\n		21.6.3 In vitro Tests\n			21.6.3.1 Cytotoxicity\n			21.6.3.2 Isoenzymes\n			21.6.3.3 Microbiological Tests\n			21.6.3.4 Mycoplasma\n	21.7 Preclinical Safety Evaluation\n		21.7.1 Acute Toxicity\n		21.7.2 Subacute Toxicity or Repetitive Doses\n		21.7.3 Chronic and Subchronic Toxicity\n		21.7.4 Teratogenicity and Reproductive Disorders\n		21.7.5 Immunotoxicity\n		21.7.6 Pharmacokinetics\n		21.7.7 Carcinogenicity or Oncogenicity\n			21.7.7.1 In vitro Preclinical Studies – Mutagenicity\n			21.7.7.2 In Vitro Mammalian Chromosomal Aberration Test (OECD 473)\n			21.7.7.3 In Vitro Mammalian Cell Gene Mutation Test (OECD 476)\n			21.7.7.4 Recombinant Bacterial Testing (OECD 471)\n	21.8 Legal Aspects for Clinical Trials\n		21.8.1 Phases of the Clinical Trial\n			21.8.1.1 Phase I\n			21.8.1.2 Phase II (Pilot Therapeutic Study)\n			21.8.1.3 Phase III\n			21.8.1.4 Phase IV\n		21.8.2 Ethical Aspects of Human Trials\n	21.9 Final Considerations\n	References\nChapter 22 Techno-Economic Evaluation of Biotechnological Processes and Pharmacoeconomic Analysis\n	22.1 Introduction\n	22.2 Bioprocess Design and Economics\n		22.2.1 Design Basis\n		22.2.2 Upstream Section\n		22.2.3 Bioreaction Section\n		22.2.4 Downstream Section\n		22.2.5 Process Flow Diagram and Process Simulation\n		22.2.6 Economic Analysis\n		22.2.7 Cost Analysis\n		22.2.8 Profitability Analysis\n	22.3 Examples of Techno-Economic Analysis\n		22.3.1 Comparison of the Insulin and BGL Production Processes\n	22.4 Beyond Techno-Economic Analysis\n	22.5 Costs Considerations in Biopharmaceuticals\n	22.6 Pharmacoeconomic Analysis\n	22.7 Biopharmaceutical Pharmacoeconomic Study Example\n	22.8 Final Considerations\n	References\nChapter 23 Perspectives for Pharmaceutical Biotechnology\n	23.1  Introduction\n	23.2  Some Neurodegenerative Diseases\n	23.3  Schistosomiasis\n	23.4  AIDS\n	23.5  Cancer\n	23.6  Stem Cells\n	23.7  Biotechnology Interfaces\n	23.8  Administration of Bioactive Molecules\n	23.9  Individualized Therapeutics\n	23.10 Synthetic Biomolecules\n	23.11 Electronics in Pharmaceutical Biotechnology\n	23.12 Final Considerations\n	References\nIndex