Download Biotechnology

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Describe the science of Biotechnology and its
product domains
List the steps to producing a GMO through use
of rDNA
Outline the steps of producing and delivering a
product made through recombinant DNA
technology
Identify the important labs and agencies in the
US, and what they’re mainly responsible for
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Apply the strategy for values clarification to
bioethical issues
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Identify products made through biotechnology
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Identify products made through biotechnology
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Insulin
Biotechnology
DNA
rDNA technology
PCR
Cloning
Fermentation
Diabetes
Antibiotics
Restriction enzyme
DNA ligase
rDNA
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Proteases
Antibodies
Pharmaceutical
R&D
Pure Science
NIH
CDC
DNA Fingerprinting
Microbial agents
Virus
Applied science
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Moral
GMO
E. coli
Plasmid
t-PA
Efficacy
Clinical trials
FDA
Cystic fibrosis
Biochemistry
Molecular biology
Genetics
ethics
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Therapeutic
EPA
USDA
Data
Hypothesis
Variable
Control
Positive control
Negative control
Concentration
Journals
Genome
bioethics
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We have been manipulating entire organisms
for thousands of years through selective
breeding to produce plant and animal products
that are
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Bigger
Healthier
More productive
Have novel phenotypes
 Scientists recently learned to manipulate not only
whole organisms, but also the molecules, cells, tissues,
and organs of which they are built
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Insulin is a protein produced in healthy
individuals that facilitates the uptake of sugar
from the blood stream into specific tissues
In some diabetic patients, the ability to produce
insulin is impaired
In the past diabetic patients had to take
injections of Bovine (cow) insulin to regulate
their blood sugar
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Problem: getting insulin from cows is not very
efficient, it’s time consuming and expensive,
and can lead to improper immune stimulation
of the recipients
Solution: Biotechnology
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We have learned to manufacture large amounts of
specific molecules, such as human insulin in bacterial
cells
 Cost effective
 Convenient
 Fast
PRESENT
PAST
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Selective breeding
Fermentation: a process
by which in an oxygen
deprived environment a
cell converts sugar into
lactic acid or ethyl
alcohol (ethanol)
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DNA technology
Recombinant DNA
(rDNA) technology
PCR
Cloning
Proteomics
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Since biotechnology is constantly changing
there is no one definitive definition
In 2003 in an international treaty on
biotechnology, modern biotechnology was
defined as :
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“the application of techniques that overcome natural
physiological reproductive or recombination barriers
and that are not techniques used in traditional
breeding and selection “ including
 In vitro nucleic acid techniques
 The fusion of cells beyond taxonomic family
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One major discovery that propelled the
biotechnology science and industry was the
discovery of restriction enzymes
Restriction enzymes: enzymes that cut DNA at
specific recognition nucleotide sequences
known as restriction sites
They were discovered in bacteria & are
thought to have evolved to provide a defense
mechanism against invading viruses
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Restriction enzymes allow us to cut and recombine
DNA molecules, often from different species
“cutting”
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DNA Ligase: an enzyme that binds the
disconnected strands of DNA together
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Reminder  DNA (deoxyribonucleic acid) is a double
stranded helical molecule that that stores genetic
information for the production of all of an organisms
proteins
“pasting”
If you put DNA into an organisms cell, that cell
will produce the protein that DNA codes for!
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Advances in rDNA technology allow
biotechnologists to manufacture a wide variety
of products either previously unavailable or
that could only otherwise be made in small
quantities
Examples:
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Insulin: for treatment of diabetes
Proteases: function in breaking down other proteins
Antibodies: proteins normally developed by the
immune system for recognizing specific molecules
called antigens on infectious agents and so limiting
their infectious ability
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PCR: Polymerase Chain Reaction is a technique
that involves copying short pieces of DNA and
using them as templates to make millions of
copies in a very short amount of time
Cloning: method of asexual reproduction that
produces identical organisms
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Done mainly in bacteria and some fungi
Only happens in animals if manipulated by scientists
to do so
Cao Yilin, a plastic surgery expert in Shanghai,
successfully reproduces a human ear on the
body of a little white mouse on Friday, April 4,
1997 in Shanghai.
Dolly, right, the first cloned sheep
produced through nuclear transfer
from differentiated adult sheep
cells, and Polly, the world's first
transgenic lamb, are in their pen at
the Roslin Institute in Edinburgh,
Scotland, in early December, 1997.
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Biotechnology is practiced in a variety of
settings , and in general the setting determine
the major emphasis
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The main difference between
Private companies: focus is making money so they
companiesthe
anddevelopment
university orand
tend to emphasize
government
labs is that
manufacture
of products
that companies
will yield high profit
must provide a product or service
margins
results
in earnings,
a nonprofit
 Thethat
do have
a Research
& Development
department,
research
facility
does that
not has potential for
but they will
only fund
research
financial pay off
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Universities & Government agencies focus on
research
 More involved in Pure science
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Pure-science: Scientific research whose main
purpose is to enrich the scientific knowledge
base
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This is the type of science nonprofit labs focus on
Applied Science: the practice of utilizing
scientific knowledge for practical purposes,
including the manufacture of a product
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This is the type of research forprofit labs focus on
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Pharmaceutical
Agricultural
Industrial
Research or production instruments, reagents,
or data
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Some companies sell their services rather than
product
Example: 23 and Me
See figure 1.13
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Research & Development : refers to the early
stages of product development that includes
discovery of the structure and function of a
potential product and the initial small scale
production
Quality Control is a measure of precision or
how well the measurement system reproduces
the same result over time and under varying
operating conditions
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National Institute of Health (NIH): the federal
agency that funds and conducts biomedical
research
Centers for Disease Control & Prevention
(CDC): National research center for developing
and applying disease prevention control,
environmental health, and health promotion
and educationactivities to promote public
health
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By the 1970’s scientists had developed new
methods including the use of restriction
enzymes and DNA ligase to create new
combinations of genetic information, or rDNA
rDNA can be inserted into cells giving these
cells a new genotype phenotype
Organisms containing these modified cells are
called Genetically Modified Organisms
(GMOs)
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GMO’s contain DNA from another organism
and will produce the new proteins encoded in
the acquired DNA
The 1st GMO’s to produce a human protein
were Escherichia coli (E. coli) bacteria
E. coli bacteria are the work horse of molecular
biology, no other bacteria is used as frequently
as this one
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Were given pieces of human DNA (genes)
containing instructions to produce a human
growth hormone called somatostatin
The somatostatin gene was carried into the E.
coli cells on tiny pieces of bacterial DNA called
Plasmids
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These recombinant DNA plasmids contained both
bacterial and human DNA
The E. coli cells read the human DNA and produced
the human protein somatostatin, which could then
be harvested and purified for use
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One of the 1st genetically engineered products
to be sold was t-PA
t-PA is a blood clot dissolving enzyme that can
be used immediately after a heart attack to
clear blocked blood vessels
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The body produces only small amounts of t-PA
To produce enough t-PA for therapeutic use
scientists genetically engineered mammalian cells
using Chinese Hamster Ovary (CHO) cells
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Ovary cells are grown in culture and
engineered with the gene that provides the
instructions to make the t-PA enzyme
By subjecting the cells to the right conditions,
the CHO cells accept and incorporate the
foreign DNA and begin the process of
transcription and translation
Large amounts of t-PA can then be extracted
and purified
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See figure 1.18
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Applications of rDNA and genetic engineering
have resulted in the creation of hundreds of
Biotech companies , specializing in all kinds of
GMOs and their protein products including
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Proteins used in pregnancy tests
Enzymes that increase the amount of juice that can
be extracted from an apple
Molecules used in vaccines
 In 2004 there were already over 370 drug products
created from rDNA techniques undergoing clinical
trials
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Strawberry plants that can grow in freezing weather
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Biotechnological advances will enable patients
to submit tissue samples and receive a
treatment regimen designed specifically for
them. This is calle Personalized medicine, or
pharmacogenomics
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pharmacogenomics: pharmaceuticals based on
genomics
Present research based on broad characteristics such
as ethnicity, gender, and family history has already
been performed and treatments have been modofied
accordingly
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The human genome project was an
international effort to sequence and map all the
DNA on the 23 pairs of human chromosomes
We know the sequence of the human genome,
but we still do not know
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all of the sequences that code for proteins
For the ones we do know about, we don’t completely
understand how, when, and where they are turned
on or off
This is called differential gene expression
DNA intro
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As sequencing human genomes becomes more
cost effective, there will come a time when
drug regimen is determined by the genotype of
the patient or the tumor being targeted
One of the 1st drugs to accomplish this is the
drug Herceptin which will only target breast
cancer cells that produce too much of the
protein HER2
The decision to administer Herceptin as part of
the chemotherapy regimen is based on a single
gene
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Some drugs, like chemotherapeutic agents,
have serious side effects because they treat the
whole organism and not just the diseased
tissue
Advances in biotechnology have enabled the
development of therapies that can be delivered
directly to diseased tissue and greatly reduce
side effects
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Rexin-G: a drug that targets pancreatic cancer is
delivered by a genetically engineered viral particle
that is designed to seek out and kill metastasized
cancer cells is in clinical trials now
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Research & Development
No matter what the product, the goal is to
make it and market it as quickly as possible
The R&D phase for a new pharmaceutical often
takes from 10-15 years for a company to move
a product through the “pipeline”
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A drug must demonstrate “proof of concept”
data in the research phase before being allowed
to move to the development phase, including
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Is it feasible to manufacture in amounts sufficient to
treat people
What needs to be done to ensure safety
What characteristics indicate it will be effective
And unfortunately, what is the profit margin
This is accomplished with a Product development
Plan
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Does the product meet the following criteria
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Does the product meet a critical need?
Is there a market large and wealthy enough to produce
enough sales
Does preliminary data support that the product will
work?
Can patent protection be secured
Can the company make a profit?
Each product in a pipeline will be reviewed
regularly and dropped from production if the
answers to these questions are unsatisfactory
See figure 1.21
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Projects then enter clinical development
Testing is done small scale before large scale
production is pursued
All pharmaceuticals have to undergo at least 3
rounds of strict clinical
trials
under the guidance of
In 2003
Pulmozyme
the FDA before being marketed
had sales of
Example: Pulmozyme $167million
is manufactured by
Genentech Inc and is an enzyme used to treat
patients with Cystic Fibrosis
People with CF experience a build up of mucus in the
lungs and intestines and is often fatal by the age 30
 Pulmozyme reduces the amount of mucus produced
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All biotechnology products have regulations
governing their production in the product
pipeline
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Environmental Protection Agency (EPA)
 The federal agency that enforces environmental laws
including the production and use of microorganisms,
herbicides, pesticides, and GMOs
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United States Department of Agriculture (USDA)
 Federal agency that regulates the use and production of
plants, plant products, plant pests, veterinary supplies
and medications, and genetically modified plants and
animals
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Being able to distinguish between right and
wrong and to make decisions
based
on
that
Also Fact: those same
knowledge is considered
having good morals
people that don’t have
Problem: not all people
have theeating
samemeat
list of
any problem
right and wrong, and soand
weusing
oftenanimal
have
different morals
products would be
to think
of
Example: Vegans don’thorrified
eat or use
any animal
homeitand
cooking
products, and manygoing
consider
immoral
to do
their pet dog Sparky or
so
their new kitten Mittens
Fact: most people eat and use
animal products
for dinner
on a daily basis without any qualms
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The study of moral standards and how they
effect conduct is called ethics
Bioethics is a term that has been coined to
describe the study of decision making as it
applies to moral decisions that need to be made
because of advances in biology, medicine, and
technology
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Many of the new biotechnologies are
controversial because they force people to
think about what they believe is right or wrong
Examples:
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Harvesting and manipulating embryonic stem cells
 It kills the human embryo
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Genetically modifying food
 Are they really safe to eat
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Prenatal genetic testing/ engineering
 Playing God
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New technologies generate ethical questions
TheBut
positions
one takes on
ethical issues
sometimes
people
are!are
based on how you feel about the knowledge
and understanding you have of a subject
There is often not a clear right or wrong answer
Most of the time you have to make a choice
between the lesser of 2 evils
So how do we decide what’s “right”
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Identify and understand the problem or issue.
List all the possible solutions to the issue
Identify the pros and cons of each of the
solutions
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Consider legal, financial, medical, personal, social,
and environmental aspects
Based on the pros and cons for each solution,
rank all the solutions from best to worst
Decide if the problem is important enough to
take a position. If it is, decide your position and
be prepared to describe and defend it
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There is no 1 correct scientific method
The scientific method is really just a series of
common sense steps and techniques that
scientists use to solve a problem or answer a
question
There are several practices used by most
scientists when conducting experimental
research
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State a testable scientific question or problem
based on some information or observation
Develop a testable hypothesis
Plan a valid experiment
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Multiple replicates
Control groups
Conduct the outlined experiment and collect
and organize data
Formulate conclusions based on experimental
data and error analysis
POSITIVE CONTROLS
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A group that will give a
predictable positive
result
NEGATIVE CONTROLS
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A group that will give a
predictable negative
result
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A good approach to writing an experimental
conclusion is to use the REE/PE/PA method
REE: Results with Evidence and Explanation
PE: Possible Errors
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Identify the sources of expreimental design errors that
would or could lead to false or misleading data, and
explain possible implications from such errors
PA: Practical Applications:
discuss the meaning or value of experimental results in
the short and long term.
 Identify how the findings could be valuable clinically, to
the company, or to the community
 Identify the direction the experiment will go take from
here
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Biotechnology: The study and manipulation of
living things or their component molecules,
cells, tissues, or organs
Product Domains
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Pharmaceutical
Agricultural
Industrial
Research or production instruments, reagents, or
data
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Identify a gene of interest
Treat gene of interest and plasmid vectors with
a restriction enzyme
Use DNA ligase to glue gene of interest to
plasmid
Insert plasmid into host organism (plant,
animal, or bacteria) through various
mechanisms
The rDNA will be used by the host organism to
produce the new protein of interest
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Come up with an Idea
Research and Development
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Proof of Concept through a Product Development
Plan
 Will it work
 Is there a market for it
 Can we make it in sufficient quantities
Large Scale Production
 Clinical Trials
 FDA approval
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NIH: the federal agency that funds and conducts
biomedical research
CDC: National research center for developing and
applying disease prevention control, environmental
health, and health promotion and education activities
to promote public health
EPA: The federal agency that enforces environmental
laws including the production and use of
microorganisms, herbicides, pesticides, and GMOs
USDA: Federal agency that regulates the use and
production of plants, plant products, plant pests,
veterinary supplies and medications, and genetically
modified plants and animals
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Identify /understand the issue
List all the possible solutions
Identify the pros and cons of each considering
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legal
financial,
Medica
personal
Social
environmental aspects
Rank solutions from best to worst
Decide and defend your position
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T-PA
Human insulin
Strawberry plants that can grow in freezing
weather
Pulmozyme