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- •Preface
- •About the Authors
- •Environmental Biotechnology
- •Microbial Biotechnology
- •Advances in Biotechnology
- •Significant Milestones in Biotechnological Research
- •Historical Contributions of some Leading Scientists
- •Contents
- •Introduction
- •Discovery of Cell and Chromosome Theory of Inheritance
- •Major Branches of Biotechnology
- •Agricultural Biotechnology
- •Medical Biotechnology
- •Pharmaceutical Biotechnology
- •Genetic Evaluation
- •Historical Perspective
- •QUESTIONS
- •Gene Cloning
- •Gene Therapy
- •Protein Biosynthesis
- •DNA Replication
- •Historical Perspectives of Plant tissue Culture
- •Introduction
- •Immune System
- •How the Immune System Works
- •Organs of Immune System
- •Lymphoid System
- •Cells of the Immune System (Fig. 2.3)
- •Principles
- •Innate Immunity
- •Adaptive Immunity
- •Types of Immunity
- •Natural Immunity
- •Acquired Immunity
- •Immune Tolerance
- •Hypersensitivity
- •Antigen
- •Antibodies
- •Haptens
- •Hapten Inhibition
- •Immunomodulators
- •Applications of Immunomodulators
- •Vaccines
- •Formulation of Vaccines
- •Standardization and Evaluation of Vaccines: WHO Approach
- •From Standards to Vaccines of Assured Quality
- •Standards for Different Stages of Vaccine Development
- •Principles for Biological Standardization
- •Vaccine Storage
- •Vaccine Storage Checklist
- •Vaccine Cold Storage
- •Introduction
- •Bioremediation
- •Ex-Situ Bioremediation
- •In-Situ Bioremediation
- •Natural Bioremediation
- •Engineered In-Situ Bioremediation
- •Management of Environmental Pollution
- •Photosynthesis
- •Biological Classification
- •Sewage Treatment by Bacteria and Algae
- •Eutrophication and Phosphorus Pollution
- •Management of Metal Pollution
- •Immobilized Cells in Management of Pollution
- •Management of Air, Waste Gases and Water Pollution
- •Bioreactors
- •Biocatalysts
- •Batch Experiments
- •Management of Water Pollution
- •Aerobic Biotreatment of Wastewater Pollution
- •Introduction
- •Proteins
- •Structure of Proteins
- •Simple proteins
- •Conjugated proteins
- •Primary Structure of Proteins
- •Secondary Structure of Proteins
- •Tertiary Structure of Proteins
- •Quaternary Structure of Proteins
- •Methods of Studying Proteins
- •Proteomics
- •Clinical Applications of Proteomics
- •Proteomics in Lab Diagnosis
- •Proteomics in the Discovery of Protein Biomarkers
- •Proteomics in Neurology
- •Proteomics in Diagnosis
- •Proteomics in the Study of Tumour Metastasis
- •Proteomics in Neurotrauma
- •Proteomics in Urological Cancer Research
- •Proteomics in the Study and Treatment of Autoimmune System
- •Proteomics in Diabetes Research
- •Proteomics in Cardiovascular Research
- •Proteomics in Investigating Plants
- •Established Cell Lines
- •Lag Phase
- •Log Phase
- •Stationary Phase
- •Primary Cell Lines
- •Interaction of Cells
- •Metabolism of Cultured Cells
- •Genetics of Cultured Cells
- •Applications of Cell Cultures
- •Mechanical
- •Biochemical
- •Animal Tissue Culture: Media Requirements
- •Culture Medium
- •Blood Plasma
- •Blood Serum
- •Preparation of Chicken Serum
- •Preparation of Mammalian Serum
- •Serum-Free Medium
- •Tissue Extracts
- •Complex Natural Medium
- •Supplemented Bovine Amniotic Fluid Medium
- •Serum-Supplemented Yeast Extract Medium
- •Serum-Supplemented Lactalbumin Hydrolysate Medium
- •Other Tissue Culture Media
- •Medium no. 612
- •Medium no. 635
- •Medium no. 858
- •Medium no. 866
- •Substrate Surfaces and Treatments
- •Disposable Plastic as a Substrate
- •Palladium and Metallic Surface as Substrates
- •Treatment of Substrate Surfaces
- •Introduction
- •Advantages and Limitations of Tissue Culture Methods
- •Primary and Established Cell Lines
- •Primary Cell Lines
- •Established Cell Lines
- •Kinetics of Cell Growth
- •Feeder Layers on Substrate
- •Gas Phase for Tissue Culture
- •Processing of Culture
- •Glassware and Apparatus
- •Reagents and Media
- •Preparation of Animal Material
- •Disaggregation of Culture
- •Physical Disruption
- •Enzymatic Digestion
- •Treatment with Chelating agents
- •Isolation of Tissue
- •Enzymatic Disaggregation
- •Mechanical Disaggregation
- •Separation of Cells
- •Somatic Cell Fusion
- •Tissue Culture
- •Primary Techniques
- •Slide or Coverslip Cultures
- •Carrel Flask Cultures
- •Test Tube Culture
- •Organ Culture
- •Culture of Embryonic Organs
- •Whole-Embryo Culture
- •Tissue Cultures in Biomedical Research
- •In Karyological Studies
- •In Identification and Hereditary Metabolic Disorders
- •In Somatic-Cell Genetics
- •In Biomedicine
- •Genetics as Applied to Medicinal Herbs
- •Mutation
- •Radiation Mutations
- •Chemical Mutations
- •Polyploidy
- •Chemodemes (Chemical Races)
- •Hybridization
- •Transgenic Plants
- •Gene Transfer
- •Transgenic Plants as Sources of Vaccines
- •Plant Tissue Cultures as Sources of Biomedicinals
- •Historical Development
- •Types of Cultures
- •Culture Medium
- •Inorganic Salts
- •Vitamins
- •Carbon Source
- •Growth Regulators
- •Organic Supplements
- •Preparation of Medium
- •Surface Sterilization of Explants
- •Establishment of Cultures
- •Callus Cultures
- •Suspension Cultures
- •Growth Measurement
- •Organogenesis
- •Phytopharmaceuticals In Plant Tissue Cultures
- •Immobilized Cell Systems
- •QUESTIONS
- •Introduction
- •Microorganisms
- •Types
- •Microbial Culture
- •Principle of Microbial Growth and Culture System
- •Culture Ingredients and Preparation
- •Types of Cultures
- •Solid or Semisolid Culture
- •Batch Culture
- •Lag Phase
- •Acceleration Phase
- •Log Phase
- •Deceleration Phase
- •Stationary Phase
- •Death Phase
- •Fed-Batch Culture
- •Variable Volume Fed-Batch
- •Advantages of Fed-Batch Reactors
- •Limitations of Fed-Batch Reactors
- •Continuous Culture
- •Homogeneously Mixed Bioreactor
- •Plug Flow Bioreactor
- •Advantage of Continuous Cultures
- •Limitation of Continuous Cultures
- •Choice of Fermenter
- •Quality Control of Microbial Culture Media
- •Raw Materials
- •Sterilization Parameter
- •Physical Parameter
- •Microbiological Parameter
- •Econometric Method
- •Productivity Ratio
- •Detection of Contamination
- •Gel Strength Parameters
- •QUESTIONS
- •Introduction
- •History of Genetics
- •Important Contributions
- •Problems and Limitations of Genetics
- •Evaluation and Molecular Biology of Plants
- •DNA as Segment of Molecular Heredity
- •Structure of DNA
- •Nucleotide
- •Double Helix
- •DNA Replication and Inheritance
- •Genes
- •Chromosomes
- •Gene Expression
- •Transcription
- •Analysis of transcription
- •Translation
- •Genetic Code
- •Mutations
- •Induced Mutation
- •Physical agents
- •Chemical agents
- •Spontaneous Mutation
- •Molecular Biotechnology
- •Techniques of Molecular Biology
- •Expression Cloning
- •Polymerase Chain Reaction
- •Gel Electrophoresis
- •Macromolecule Blotting and Probing
- •QUESTIONS
- •Introduction
- •Outline of Recombinant DNA Technology
- •Principle of Gene Cloning
- •Genomic DNA Libraries
- •Construction
- •Preparation
- •Isolation of Gene Sequence from the Libraries
- •Plasmid as Cloning Vector
- •Role of DNA Ligase
- •Applications of Recombinant Dna Technology
- •Production of Biologicals
- •Insulin
- •Production of Interferons (Fig. 9.3)
- •Growth Hormones
- •Erythropoietin
- •Cytokines
- •Interleukins
- •Tumour Necrosis Factor and Lymphotoxin
- •Vaccines
- •Live vaccines
- •Killed vaccines
- •Production of Human Follicle-Stimulating Hormone
- •Disease Diagnosis by Recombinant Dna Methods
- •Applications in the Pulp and Paper Industry
- •Applications in Antibiotic-Producing Microorganisms
- •QUESTIONS
- •Introduction
- •Transformation: Discovery and Types
- •Natural Transformation
- •Artificial Transformation
- •Types of Reactions Mediated by Microorganisms
- •Oxidative Biotransformations
- •Hydrolysis of Oxiranes, Nitriles and Amides
- •Design for Biotransformation
- •Biotransformation of Steroids
- •Microbial Transformation of Triterpenoids
- •Biotransformation of Pentacyclic Triterpenoids
- •Microbial Transformation of Cholesterol by Mycobacterium Smegmatis
- •Microbial Transformation of Progesterone in Vitro
- •QUESTIONS
- •Introduction
- •Principle for Creation of Hybridoma Cells
- •Immunization
- •Cell Fusion
- •Formation and Selection of Hybrid Cells
- •Screening of Products
- •Cloning and Propagation
- •Characterization and Storage
- •Large-Scale Production of Monoclonal Antibodies
- •Human mAbs
- •Advantages and Limitations of mAbs
- •Advantages
- •Limitations
- •Fusion and Culture of Hybridomas
- •Cloning of Preservation of Hybridomas
- •Applications of Monoclonal Antibodies
- •Diagnostic Applications
- •Investigational and Analytical Applications
- •Identification and Isolation of Lymphocytes Phenotyping
- •Purification of Proteins
- •Miscellaneous Applications
- •Application of Catalytic mAbs
- •mAbs in Drug Targeting
- •QUESTIONS
- •Historical Development of Antibiotics
- •Discovery of Penicillin and Other Antibiotics
- •Antimicrobial Spectrum
- •Standardization of Antibiotics
- •Minimal Inhibitory Concentration (MIC)
- •Agar Diffusion Tests
- •MIC Determination by Broth Dilution (Using Sensititre)
- •Screening of Soil for Organisms Producing Antibiotics
- •Isolation of Microorganisms
- •Fermenter or Bioreactor
- •Types of End Products of Fermentation
- •Design of Industrial Fermentation Process
- •Solid-State Fermentation
- •Submerged Fermentation
- •Batch Fermentation
- •Other Fermenters
- •Fermenter Design and Control
- •Isolation of Mutants
- •Factors Influencing Rate of Mutation
- •Isolation of Fermentation Products
- •Production of Penicillin
- •Bioproduction of Streptomycin
- •Bioproduction
- •Tetracycline Production with Sweet Potato Residue by SSF
- •Materials and Methods
- •QUESTIONS
- •Introduction
- •Methods of Gene Transfer in Plants
- •Vector-Mediated Gene Transfer
- •Vector-Free DNA-Mediated (Direct) Gene Transfer
- •Physical Method
- •Chemical Method
- •Vector-Mediated Gene Transfer
- •Using Vector of Agrobacterium
- •Vector-Free DNA-Mediated (Direct) Gene Transfer
- •Physical Methods
- •Electroporation
- •Microinjection
- •Macroinjection
- •Biolistics or Microprojectiles for DNA Transfer
- •Liposome-Mediated Gene Transfer
- •Some Other Techniques Used for Liposomes
- •Chemical Methods
- •PEG-Mediated Transfection
- •Calcium Phosphate-Mediated DNA Transfer
- •Transfer of DNA by Polycation-DMSO
- •DNA Transfer by DEAE-D
- •Use of Polybrene
- •Gene Transfer through Peptide
- •Horizontal Gene Transfer Technology
- •Transformation
- •Mechanism of Transformation
- •Transduction
- •Conjugation
- •Mechanism of Conjugation
- •Specialized Transduction
- •HGT in Prokaryotes
- •Protoplast Fusion
- •Enzymes Used for Breaking of Cell Walls
- •Methods of Protoplast Fusion
- •Protoplast Fusion in Plants
- •Protoplast Fusion in Fungi
- •Protoplast Technology for Streptomyces spp.
- •Protoplast Fusion in Bacteria
- •Gene Cloning
- •Cloning
- •Gene Targeting
- •Stem Cells
- •Development of Hybridoma by Monoclonal Antibodies
- •Hybridoma Technology
- •Hybridomas
- •Hybridization Techniques
- •Southern Blot
- •Northern Blot
- •Western Blot
- •Some Important Medicines Produced by Biotechnology
- •Humulin
- •Activase
- •Humatrope
- •Hepatitis B Vaccine (Recombinant)
- •Gene Transfer in Humans
- •Techniques of Gene Transfer in Human Cells
- •Adenoviruses
- •Applications of Gene Transfer
- •Role of Gene Transfer in Human Diseases
- •Gene Transfer Technology in Other Areas
- •QUESTIONS
- •Introduction
- •Public Health
- •Basic Sciences of Medicine
- •Edible Vaccines
- •Important Steps in the Production of Edible Vaccines
- •Challenges in the Production of Edible Vaccines
- •Nonscientific Challenge
- •Regulatory Issues
- •Clinical Trials
- •Some Edible Vaccines Produced by Plants
- •Advantages and Limitations of Edible Vaccines
- •QUESTIONS
- •Introduction
- •Production of Antibiotics
- •Production of Vaccines
- •New Generation of Vaccines
- •Human Therapeutics from Recombinant DNA Technology
- •Production in Lactic Acid
- •Plant Tissue Culture for Production of Plant Secondary Metabolites
- •Applications of Biotechnology in the Diagnosis of Diseases
- •Applications of Biotechnology in Agriculture
- •Reduced Vulnerability of Crops to Environmental Stresses
- •Reduced Dependence on Fertilizers, Pesticides and Other Agrochemicals
- •Production of Novel Substances in Crop Plants
- •Applications of Ultrasound to Biotechnology
- •Testing of Pharmaceuticals
- •QUESTIONS
- •Introduction
- •Natural Product Drug Discovery
- •Screening of Natural Products
- •Structural Elucidation
- •QUESTIONS
- •Introduction
- •History of Enzymes
- •Enzyme Structure
- •Enzyme Kinetic
- •Importance of Enzyme Kinetics
- •Factors Affecting Enzyme Reaction Velocity
- •Bioproduction of Enzymes
- •Microorganisms
- •Enzyme Properties
- •Enzyme Production
- •Enzyme Immobilization
- •Advantages of Immobilized Enzymes
- •Techniques of Enzyme Immobilization (Fig. 17.4)
- •Physical Immobilization
- •Chemical Immobilization
- •Adsorption
- •Cell Immobilization
- •Immobilization of Plant Cells
- •Method of Immobilization
- •Immobilized Plant Cell Reactors
- •Immobilization of Bacterial Cell
- •Procedural Details
- •Immobilized Cell Preparation
- •Immobilized Cell Reactor Construction and Operation
- •Some Important Enzymes
- •Amylase
- •Hyaluronidases
- •Protease
- •Streptokinase
- •Penicillinase
- •Enzyme Engineering
- •Applications of Enzymes
- •Biosensors
- •Types of Biosensors
- •Applications of Biosensors
- •QUESTIONS
- •Introduction
- •Computational Gene
- •Application in Therapy of Cancer
- •Challenges
- •Nanolithography
- •Optical Lithography
- •Other Nanolithography Techniques
- •Bottom-up Methods
- •Nanomedicine and Nanodevices
- •Medical Uses of Nanomaterials
- •In Drug Delivery
- •Protein and Peptide Delivery
- •Neuroelectronic Interfaces
- •In Cancer
- •In Surgery
- •Medical Applications of Molecular Nanotechnology
- •Nanorobots
- •Cell Repair Machines
- •Nanonephrology
- •Nanosensor
- •Predicted Applications
- •Existing Nanosensors
- •Production Methods
- •Economic Impact
- •Social Impact
- •Nanoparticles
- •Background
- •Properties
- •Synthesis
- •Colloids
- •Morphology
- •Characterization
- •Functionalization
- •Safety
- •Synthetic Biology
- •QUESTIONS
- •Introduction
- •Biopharmaceuticals of Animal Origin
- •Sex Hormones
- •Androgens
- •Oestrogen
- •Progesterones and Progestogens
- •Biopharmaceuticals of Plant Origin
- •Recombinant Proteins
- •Plant-Derived Therapeutics
- •Alternative Plant-Based Expression System
- •Pharmaceutical Substances of Microbial Origin
- •Major Families of Antibiotics
- •Recent Advancs in Biopharmaceuticals
- •Applications of Carrier Erythrocytes in the Delivery of Biopharmaceuticals
- •Important Features of Carrier Erythrocytes
- •Research Advances on Transgenic Plant Vaccines
- •Recent Advances in Analysis of Peptides and Proteins by Mass Spectrometry
- •Advances in Chloroplast Engineering
- •QUESTIONS
- •Appendix
- •Glossary
- •Bibliography
- •Index

Contents
CHAPTER 1: Historical Perspectives and Milestones ...................................................................... 1
Introduction ............................................................................................................................................... 1
Discovery of Cell and Chromosome Theory of Inheritance ........................................................................ 1
Major Branches of Biotechnology .............................................................................................................. 2
Advances in Biotechnology .......................................................................................................................4
Signifi cant Milestones in Biotechnological Research ................................................................................. 4
Historical Contributions of Some Leading Scientists ................................................................................. 4
Genetic Evaluation ................................................................................................................................... 10
Historical Perspectives of Plant Tissue Culture .......................................................................................13
Questions ................................................................................................................................................ 14
CHAPTER 2: Immunology and Immunological Preparations ........................................................ 15
Introduction ............................................................................................................................................. 15
How the Immune System Works ............................................................................................................. 16
Principles ............................................................................................................................................... 19
Types of Immunity .................................................................................................................................. 21
Immune Tolerance ................................................................................................................................... 22
Hypersensitivity ....................................................................................................................................... 23
Antigen–Antibody Reactions and Their Applications ............................................................................... 24
Immunomodulators ................................................................................................................................. 26
Applications of Immunomodulators ........................................................................................................ 29
Vaccines ..................................................................................................................................................30
Questions ................................................................................................................................................ 34
CHAPTER 3: Environmental Biotechnology .................................................................................. 35
Introduction ............................................................................................................................................. 35
Bioremediation ........................................................................................................................................ 36
Management of Environmental Pollution ................................................................................................. 36
Management of Air, Waste Gases and Water Pollution ............................................................................ 40
Questions ................................................................................................................................................ 42

Contents
x
CHAPTER 4: Proteins and Proteomics .......................................................................................... 43
Introduction ............................................................................................................................................. 43
Proteins ................................................................................................................................................... 44
Proteomics .............................................................................................................................................. 50
Questions ................................................................................................................................................ 53
CHAPTER 5: Animal Cell and Tissue Biotechnology ..................................................................... 55
Introduction ............................................................................................................................................. 55
Primary and Established Cell Lines .........................................................................................................56
Kinetics of Cell Growth ............................................................................................................................ 57
Animal Tissue Culture: Media Requirements ........................................................................................... 60
Questions ................................................................................................................................................ 73
CHAPTER 6: Medicinal Plant Biotechnology ................................................................................ 75
Genetics as Applied to Medicinal Herbs ..................................................................................................75
Mutation .................................................................................................................................................. 77
Polyploidy ............................................................................................................................................... 79
Chemodemes (Chemical Races) .............................................................................................................. 81
Hybridization ........................................................................................................................................... 81
Genetic Engineering and Recombinant DNA Technology in Plants .......................................................... 82
Plant Tissue Cultures as Sources of Biomedicinal Compounds ............................................................... 86
Historical Development ..........................................................................................................................88
Types of Cultures ..................................................................................................................................... 89
Culture Medium ....................................................................................................................................... 90
Surface Sterilization of Explants .............................................................................................................. 91
Establishment of Cultures .......................................................................................................................92
Phytopharmaceuticals in Plant Tissue Cultures ....................................................................................... 94
Bioproduction of Useful Metabolites in Hairy Root and Multiple Shoot Cultures ..................................... 95
Questions ................................................................................................................................................ 97
CHAPTER 7: Microbial Culture and Fermentation Process........................................................... 99
Introduction ............................................................................................................................................. 99
Microorganisms .................................................................................................................................... 100
Microbial Culture .................................................................................................................................. 100
Quality Control of Microbial Culture Media ............................................................................................ 107
Questions .............................................................................................................................................. 109
CHAPTER 8: Genetics and Molecular Biotechnology ................................................................. 111
Introduction ........................................................................................................................................... 111
DNA as Segment of Molecular Heredity ................................................................................................113
Genes .................................................................................................................................................... 116
Mutations .............................................................................................................................................. 123

Historical Perspectives and Milestones in Pharmaceutical Biotechnology
xi
Contents
Molecular Biotechnology ....................................................................................................................... 124
Questions .............................................................................................................................................. 127
CHAPTER 9: Recombinant DNA Technology ............................................................................. 129
Introduction ........................................................................................................................................... 129
Outline of Recombinant DNA Technology .............................................................................................. 130
Introduction of the Recombinant DNAs into a Suitable Host Cell and Their Identifi cation ..................... 132
Applications of Recombinant DNA Technology...................................................................................... 134
Production of Human Follicle-Stimulating Hormone ............................................................................. 139
Disease Diagnosis by Recombinant DNA Methods ................................................................................ 139
Recombinant DNA Technology for Improving Milk Production ............................................................. 140
Applications in the Pulp and Paper Industry ......................................................................................... 140
Applications in Antibiotic-Producing Microorganisms ..........................................................................140
Genetic Engineering as a Part of Recombinant DNA Technology ........................................................... 141
Questions .............................................................................................................................................. 142
CHAPTER 10: Microbial Transformation .................................................................................... 143
Introduction ........................................................................................................................................... 143
Types of Reactions Mediated by Microorganisms ................................................................................. 145
Design for Biotransformation ................................................................................................................ 149
Biotransformation Process and Its Improvement with Special Reference to Steroids ........................... 150
Questions .............................................................................................................................................. 153
CHAPTER 11: Hybridoma Technology and Monoclonal Antibodies ........................................... 155
Introduction ........................................................................................................................................... 155
Principle for Creation of Hybridoma Cells .............................................................................................. 156
Large-Scale Production of Monoclonal Antibodies ................................................................................ 158
Applications of Monoclonal Antibodies .................................................................................................160
Questions .............................................................................................................................................. 165
CHAPTER 12: Antibiotics ........................................................................................................... 167
Historical Development of Antibiotics .................................................................................................... 167
Antimicrobial Spectrum ......................................................................................................................... 171
Standardization of Antibiotics ................................................................................................................ 172
Screening of Soil for Organisms Producing Antibiotics ......................................................................... 174
Fermenter or Bioreactor ........................................................................................................................176
Design of Industrial Fermentation Process ............................................................................................ 178
Fermenter Design and Control ............................................................................................................... 180
Isolation of Mutants ..............................................................................................................................181
Factors Infl uencing Rate of Mutation ..................................................................................................... 182
Isolation of Fermentation Products .......................................................................................................183
Questions .............................................................................................................................................. 187
xi

xii
Contents
CHAPTER 13: Genetic Recombination ....................................................................................... 189
Introduction ........................................................................................................................................... 189
Methods of Gene Transfer in Plants....................................................................................................... 190
Horizontal Gene Transfer Technology ...................................................................................................200
Protoplast Fusion .................................................................................................................................. 205
Gene Cloning ......................................................................................................................................... 207
Development of Hybridoma by Monoclonal Antibodies ......................................................................... 208
Some Important Medicines Produced by Biotechnology ....................................................................... 211
Gene Transfer in Humans ...................................................................................................................... 213
Applications of Gene Transfer ................................................................................................................ 213
Questions .............................................................................................................................................. 216
CHAPTER 14: Medicine and Edible Vaccines ............................................................................. 217
Introduction ........................................................................................................................................... 217
Edible Vaccines ..................................................................................................................................... 219
Questions .............................................................................................................................................. 225
CHAPTER 15: Broad Applications of Pharmaceutical Biotechnology ......................................... 227
Introduction ........................................................................................................................................... 227
Applications of Biotechnology in the Production of Biomolecules ......................................................... 228
Applications of Biotechnology in the Diagnosis of Diseases .................................................................. 231
Applications of Biotechnology in Agriculture ......................................................................................... 231
Applications of Ultrasound in Biotechnology ......................................................................................... 232
Testing of Pharmaceuticals.................................................................................................................... 232
Questions .............................................................................................................................................. 232
CHAPTER 16: Biotechnology in Drug Discovery ........................................................................ 233
Introduction ........................................................................................................................................... 233
Current Trends in Modern Pharmaceutical Analysis for Drug Discovery ............................................... 235
Recent Pharmacokinetic Advances in Drug Discovery and Development .............................................. 235
Natural Product Drug Discovery ............................................................................................................ 235
Questions .............................................................................................................................................. 237
CHAPTER 17: Immobilization of Enzymes ................................................................................. 239
Introduction ........................................................................................................................................... 239
History of Enzymes ...............................................................................................................................240
Enzyme Structure .................................................................................................................................. 241
Enzyme Kinetic ...................................................................................................................................... 242
Bioproduction of Enzymes ..................................................................................................................... 246
Enzyme Production ............................................................................................................................... 248
Enzyme Immobilization ......................................................................................................................... 249
Immobilization of Plant Cells ................................................................................................................. 255

Contents
xiii
Enzyme Engineering ..............................................................................................................................
259
Applications of Enzymes .......................................................................................................................260
Biosensors ............................................................................................................................................ 262
Questions .............................................................................................................................................. 267
CHAPTER 18: Nanobiotechnology ............................................................................................. 269
Introduction ........................................................................................................................................... 269
Computational Gene .............................................................................................................................. 270
Nanolithography .................................................................................................................................... 272
Nanomedicine and Nanodevices ............................................................................................................ 274
Medical Applications of Molecular Nanotechnology .............................................................................. 277
Questions .............................................................................................................................................. 286
CHAPTER 19: Recent Advances in Biopharmaceuticals ............................................................. 287
Introduction ........................................................................................................................................... 287
Advantages of DNA Technology in the Production of Biopharmaceuticals ............................................ 288
Biomolecules Produced Through Recombinant DNA Technology ......................................................... 289
Biopharmaceuticals of Animal Origin ....................................................................................................290
Biopharmaceuticals of Plant Origin .......................................................................................................292
Pharmaceutical Substances of Microbial Origin .................................................................................... 293
Recent Advances in Biopharmaceuticals ............................................................................................... 300
Questions .............................................................................................................................................. 303
Appendix .................................................................................................................................... 305
Glossary ...................................................................................................................................... 313
Bibliography ............................................................................................................................... 321
Index .......................................................................................................................................... 327

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INTRODUCTION
The term biotechnology represents a fusion or an alliance between biology and technology. It is
a newly discovered discipline for old-age practices. Biotechnology, as an integral component of
biosciences, has derived its strength from different disciplines of science and technology. It is an
interdisciplinary pursuit with multidisciplinary applications.
DISCOVERY OF CELL AND CHROMOSOME THEORY OF INHERITANCE
Robert Hooke is credited with the discovery of cell. An important discovery made by Robert Brown
(1831) was the presence of a small sphere within the cells of orchid roots. In 1839, Hugo von Mohl
and J. Purkinje named the jelly-like substance as protoplast. In 1885, Virchow explained that cells
are derived from pre-existing cells.
In 1902, cytologists Walter Sutton and Theodor Boveri independently came to the conclusion
that the behaviour of chromosomes at meiosis can serve as the cellular basis of both segregation
and independent assortment. The theory of heredity of chromosomes was further expanded by
Thomas H. Morgan on fruit fly drosophila. The term factor as the basic unit of inheritance was
replaced with the term gene by Johannsen in 1909. The term mutation was coined in 1901 by Hugo
de Vries to explain the variation he observed in the plant Oenothera lamarckiana (primrose).
Historical Perspectives and
Milestones
1
CHAPTER
Introduction
Discovery of Cell and Chromosome
Theory of Inheritance
Major Branches of Biotechnology
Advances in Biotechnology
Signifi cant Milestones in Biotechnological
Research
Historical Contributions of Some
Leading Scientists
Genetic Evaluation
Historical Perspectives of Plant
Tissue Culture
Questions

Textbook of Pharmaceutical Biotechnology
2
The origin of biotechnology dates back to 6000 BC, the same year when the yeast was first
used to produce beer and wine. The history of biotechnology begins with zymotechnology, which
commenced with a focus on brewing techniques for beer. By World War I, zymotechnology
expanded to cover larger industrial issues, and the potential of industrial fermentation gave rise
to biotechnology. Yoghurt was produced from bacteria. Louis Pasteur, also known as the father
of biotechnology, identified the role of microorganisms in fermentation. The biotechnological
revolution began in the late 1970s and early 1980s when scientists understood the genetic
constitution of living organisms. Biotechnology, as an applied bioscience, has been effectively
utilized for better industrial therapeutic production. As a matter of fact, every discipline of science
has contributed either directly or indirectly to the growth of biotechnology (Fig. 1.1).
MAJOR BRANCHES OF BIOTECHNOLOGY
Biotechnology is the application of scientific and engineering principles to the processing of
materials by biological agents—microorganisms, plants and animal cells. Biotechnology can be
represented as a mixture of various biological sciences (biosciences) for better service in the field
of pharmaceuticals (Fig. 1.2).
Agricultural Biotechnology
It deals with a group of scientific techniques that are used to create, improve or modify plants
and animals. It encompasses the knowledge of biosciences concerned with plants and agriculture,
such as plant tissue culture, production of haploid plants, somaclonal variation, micropropagation
cryopreservation methodology in the genetic engineering of plants, application of transgenic
plants, etc.
Figure 1.1 Basic strength for advancement of biotechnology.
Biochemistry
Biophysics
Biotechnology
Genetics
Cell biology
Microbiology
Engineering
biotechnology

Historical Perspectives and Milestones
3
Forensic Biotechnology
It is the method for detecting unique DNA pattern of organism, and it includes DNA
fingerprinting.
Medical Biotechnology
It deals with the utilization diagnostic kits for the detection of different diseases. It has wide
applications in human health care, and it includes information related to gene therapy, DNA in
disease diagnosis, fingerprinting, specialized products of DNA biotechnology, etc.
Pharmaceutical Biotechnology
It is a major branch of biotechnology, which includes production of therapeutic proteins and
hormones, fermentation products such as antibiotics, vaccines and drugs, etc.
Environmental Biotechnology
It encompasses the information on biotechnology with references to environment such as
environment pollution, treatment modalities, etc.
Microbial Biotechnology
The information pertaining to fermentation technology, downstream processing, enzyme
technology, microbial production of organic solvents, organic acids, antibiotics, amino acids,
Biotechnology
Agricultural
biotechnology
Environmental
biotechnology
Animal cell
biotechnology
Microbial
biotechnology
Medical
biotechnology
Forensic
biotechnology
Pharmaceutical
biotechnology
Molecular
biotechnology
Genetic
biotechnology
Figure 1.2 Branches of biotechnology.

Textbook of Pharmaceutical Biotechnology
4
vitamins, foods and beverages, polysaccharides, biomass–bioenergy, etc., is covered under
microbial biotechnology.
Biotechnology has several other branches also, with varied applications, such as molecular
biotechnology, aquatic biotechnology, animal biotechnology, environmental biotechnology, leather
biotechnology, mining and metal biotechnology, textile biotechnology, genetic engineering, etc.
ADVANCES IN BIOTECHNOLOGY
With the advent of information technology, modern technologies give rise to genomics, proteomics
and cellomics that promise to develop into the technology with applications in medicine, food,
agriculture, etc. The present focus of biotechnology is centred basically on genomics and
postgenomic. Since 2001, different methods have been developed enabling rapid sequencing of
more than 50 microbial, plant, animal and human genomes.
Newer frontiers in biotechnology include development of nucleic acid probes, production of
monoclonal antibodies and molecular markers. Development of microbial inoculants covering
biopesticides, biofertilizers and genetic modifications of microbes through DNA recombination
are other important applications. Novel approaches such as proteomics and structure biology are
contributing to the understanding of chemistry of life and disease.
Different techniques of population genetics and biometric analysis are complemented with
modern methods in reproductive biology and gene diagnosis. Artificial insemination, in vitro
fertilization and embryo transfer are important applications of biotechnology in animal breeding.
SIGNIFICANT MILESTONES IN BIOTECHNOLOGICAL RESEARCH
During the 20th century, the pharmaceutical industry witnessed a series of developments in
science and technology, which had generated new opportunities for biotechnological discoveries.
The important historical developments in biotechnology, recent milestones in biotechnology
and chronological sequences of important biotechnological products (drugs, vaccines and other
therapeutics) are listed in Tables 1.1, 1.2 and 1.3, respectively.
HISTORICAL CONTRIBUTIONS OF SOME LEADING SCIENTISTS
1. Antony van Leeuwenhoek (1632–1723) is the greatest of all microscopists. He was the first
to observe and accurately describe the shape of human red blood cells. He observed and
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