Biotechnology and the Workplace

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22 Φεβ 2013 (πριν από 4 χρόνια και 5 μήνες)

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Chapter 1

Biotechnology and


the Workplace

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

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

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

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

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)


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

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

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


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

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

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

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
α
-
2b
-
1986


Treats variety of viral diseases and cancers


Hairy cell leukemia, AIDS
-
related Kaposi’s 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

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

The
plasmid

is digested with
Eco
RI

, 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
.

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 rbc’s


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

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

Other Applications


Bioremediation


Clean
-
up of contaminated soil and water


Identification of individuals


Human DNA sequences very distinct (except for
identical twins)


DNA fingerprinting

The Organization of a Biotechnology
Company


Research and Development
-

R&D


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

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 lots’o this)

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

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

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


R&D 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

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