Biotechnology and the Workplace

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Chapter 1

• Biotechnology and the Workplace

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Introduction

• Biotechnology
• The use of organisms/materials derived from
  organisms
• Make useful products
• Not a recent phenomenon
• Used organisms to make products
• Wine, cheese, bread
• Usually referring to something more dazzling
• Cloned sheep, gene therapies, DNA fingerprints

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Introduction

• Modern biotechnology
• Rooted in basic laboratory research
• Broadens/deepens understanding of how living
  organisms work
• The application of knowledge useful to humans
• Knowledge gained applied to create modern
  biotechnology

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Technologies of Modern Biotechnology

• Modern biotechnology
• Manipulation of DNA
• Tools cut DNA at specific sites
• Enzymes to splice DNA strands together
• Techniques to visualize DNA
• Techniques to separate DNA fragments from one
  another
• Techniques to identify fragments of DNA with
  SPECIFIC sequences
• Techniques to amplify DNA
• Techniques to sequence and synthesize DNA

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Technologies of Modern Biotechnology

• Tools and the terms
• Allow scientists to introduce genetic
  information/DNA from one organism to another
• Genetically modified/genetically engineered
• When a biologist causes a cell or organism to
  take up a gene from another organism

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Technologies of Modern Biotechnology

• More terms
• Recombinant DNA
• DNA that contains sequences of DNA from different
  sources that were brought together using the
  tools of biotechnology
• Often referred to as revolutionary
• There is POWER in the manipulation of DNA!
• DNA directs the cell in making a specific protein
• Called expression (the protein is expressed)

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Technologies of Modern Biotechnology

• How is DNA transferred?
• Plasmids
• Plasmids are molecules of DNA that are found in
  bacteria separate from the bacterial chromosome.
  They
• are small (a few thousand base pairs)
• usually carry only one or a few genes
• are circular
• have a single origin of replication
• Act as a vector
• Carries a desired gene into the recipient cell
• Other vectors include viral vectors and direct
  injection
• Under proper conditions, plasmids are readily
  taken up by recipient bacterial cells

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Technologies of Modern Biotechnology

• After bacteria transformed, then what?
• Fermentation
• The large-scale cultivation of microorganisms
• Alter the bacteria
• Grow in media
• Harvest the protein of interest
• What about other types of cells?
• Possible to introduce gene of interest into
  cultured mammalian cells/other multicellular
  organisms
• Cultured cells
• Cells grown in flasks, dishes, vats, or other
  containers outside a living organism
• Grow more slowly than bacteria
• More fragile
• More complex nutrient requirements
• Introducing gene of interest into plants and
  animals
• Transgenic
• A plant or animal whose cells are genetically
  modified using the techniques of biotechnology

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Technologies of Modern Biotechnology

• Genes responsible for particular traits or
  disease susceptibility are chosen and extracted.
• Next they are injected into fertilized mouse
  eggs.
• Embryos are implanted in the uterus of a
  surrogate mother.
• The selected genes will be expressed by some of
  the offspring

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Technologies of Modern Biotechnology

• What about humans?
• Raises critically important societal and ethical
  issues
• Can it be passed to offspring
• Present research into curing illness
• Intended to cure individuals
• Not affect children of the treated person, just
  the illness or condition
• What use then?
• Explore role of genes
• Mechanisms of control

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Technologies of Modern Biotechnology

• Other uses proving to be beneficial to humans
• Monoclonal antibodies
• Antibodies
• Proteins of the immune system
• Recognize/bind to substances invading the body
• Bind only to a specific target
• Exceptionally homogeneous populations of
  antibodies directed against a specific target
• Produced by hydridoma cells
• Result from the fusion of an antibody-production
  cell and a cultured cell
• Very specific and useful
• Example home pregnancy tests

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Applications-Medical/Veterinary

• Biopharmaceuticals
• Pharmaceutical products, like insulin or Factor
  VIII, that are manufactured using genetically
  modified organisms as production systems
• Transgenic animals
• Can produce desired protein
• Typically secreted into milk
• Isolated and purified
• Transgenic plants
• Genes introduced into crop plants
• Cultivated and harvested to obtain product of
  interest
• Gene therapy
• Involves replacing a gene that is missing, or
  correcting the function of a faulty gene, in
  order to treat or cure an illness

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Examples-E. coli

• Recombinant DNA products produced in E. coli
• Human insulin-1982
• Before purified from pancreas glands of animals
  for human consumption
• Genetically modified
• More reliable source
• Less likely to cause allergic reactions
• Human Growth Hormone-1985
• Before, dwarfism treated with hormone purified
  from pituitary glands from cadavers
• Some using this died from neurodegenerative
  disease-Creutzfeldt-Jakob disease!!!!!!
• Interferon a-2b-1986
• Treats variety of viral diseases and cancers
• Hairy cell leukemia, AIDS-related Kaposis
  sarcoma, renal cell carcinoma, and chronic
  hepatitis B
• Known about in 1957, but difficult to isolate
  (viral disease in patients, ONLY)
• Not available until recombinant methods developed

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Examples- Yeast Cells

• Recombinant DNA products produced in yeast cells
• Hepatitis B vaccine-1986
• Viral illness-no treatment currently available
• Before recombinant methods, vaccine prepared from
  plasma of hepatitis-infected humans
• Limited sources
• Concerns about purity
• From yeast cells
• Unlimited sources
• Reduced possibility of contamination

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The plasmid is digested with EcoRI , which cuts
it at the unique G/AATTC site in the polylinker
region, producing two "sticky" single-stranded
TTAA-3' overhanging ends. Foreign DNA is
digested with the same enzyme, which produces a
large series of fragments, all with the same
ends. These sticky ends allow one of the foreign
DNA fragments to be inserted into the plasmid
polylinker the corresponding sticky ends anneal
to each other. The gaps are closed with DNA
ligase, producing a recombinant plasmid.
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Examples- Mammalian cells in culture

• Recombinant DNA products produced in mammalian
  cells in culture
• Erythropoietin-1989
• Used to treat anemia from renal failure or AZT
  treatment
• EPO produced in the kidney
• Stimulates production/maturation of rbcs
• Found in small quantities in urine
• Never available before recombinant methods
• Anemia from kidney disease treated with
  transfusions
• Factor VIII
• Treatment for hemophilia
• Before recombinant methods, source was human
  plasma
• Patients contracted AIDS

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Applications-Agricultural/Food Related

• Breeding programs for plants and animals
• Enhance characteristics
• Benefit from biotechnological methods
• Very specific genetic manipulation organisms
• Does away with most undesirable traits passed on
  by traditional manipulations
• VERY controversial
• Not as safe as conventional foods?
• Introduction of genetically modified organisms
  into environment may have unforeseen and adverse
  effects
• Benefits
• End severe food shortages
• Enhance nutrition of plants and animals
• Reduce reliance on chemical pesticides

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Other Applications

• Bioremediation
• Clean-up of contaminated soil and water
• Identification of individuals
• Human DNA sequences very distinct (except for
  identical twins)
• DNA fingerprinting

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The Organization of a Biotechnology Company

• Research and Development- RD
• Find a potential product WITH commercial value
• Performs research relating to the potential
  product
• Characterizes properties of the product
• Composition, physical and chemical properties
• Strength, potency, or effect of the product
• Purity of product
• Steps required to avoid contamination
• Stability/shelf-life of product
• Applications of product
• Safety concerns in use of the product
• Establishes product specification
• Properties that every batch of final product must
  have to be released for sale
• Develop testing methods to ensure meets
  specifications
• Develop processes to make product
• Describe cells or microorganisms to make product
• Raw materials needed and specifications of those
  materials
• Equipment needed
• Plan for production

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The Organization of a Biotechnology Company

• Production Unit
• Make product
• Scale-up
• Work with large-scale equipment/large volume
  reactions
• Monitor and control environment as required for
  the product
• Proper temperature, sterility, etc.
• Cleaning, calibration, and maintenance of
  equipment
• Follow written procedures associated with
  producing product
• Monitor processes
• Initiate corrective actions if problems arise
• Complete paperwork!!!!!! (ALWAYS lotso this)

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The Organization of a Biotechnology Company

• QA/QC
• Monitor equipment, facilities, environment,
  personnel, and product
• Reviewing all production procedures used in the
  company
• Ensure accuracy of all documents
• Test samples of product and materials need for
  producing product
• Compare data to established standards
• Decide whether or not to approve product for
  release
• Review customer complaints

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The Organization of a Biotechnology Company

• Other functions
• Engineering or facilities management
• Ensures systems that control building are
  operating properly
• Large equipment properly installed and
  functioning
• Facility maintenance and housekeeping
• Receiving and shipping
• Dispensing
• Metrology
• Marketing and sales

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Different Types of Biotechnology Work Environments

• Basic Biological Research Labs in Academic or
  Government
• Basic research
• Investigate fundamental problems in biology
• Product is knowledge or information
• May result in a commercial product
• RD laboratories associated with industry
• Investigate questions in biology intended to
  result in commercial products
• Production facilities
• Make products
• Usually use large scale equipment
• Testing laboratories
• Analytical, quality control, forensic,
  microbiology, metrology, clinical testing labs

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Different Types of Biotechnology Work Environments

• Regardless of work environment
• Scientists and technicians involved in the
  physical aspects of acquiring data or making a
  product
• Grow bacterial cells
• Plants
• Animals
• Work with materials derived from organisms
• Make measurements, prepare reagents,
  gather/record data