Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.us
Pharmaceutical Biotechnology
Lecture 21
Part Ib. Emergent Technologies:
DNA-Based Biotechnology and
Pharmaceutical Drug Development
© life_edu
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
Kimberly Nelson
OnCampus Live
BCH 190, MIC 190, AFS 190, NRS 190, PLS 190
OnLine BCH 190
A Sweeping General Survey on Life and Biotechnology
A Public Access College Course
The University of Rhode Island
Issues in Biotechnology:
Biotechnology, Our Society and Our Future
life
edu.us
Issues in Biotechnology:
The Way We Work With Life
Dr. Albert P. Kausch
life edu.us
BCH 190
Section II.
The Applications of
Biotechnology
A Sweeping General Survey on Life and Biotechnology
© life_edu
The University
of Rhode Island
Pharmaceutical Biotechnology
What is it?
• Where do our Medicines come from?
• History
• Alternative Therapies and Science
• How is DNA-based biotechnology used in current
pharmaceutical drug development improvement?
• Small Molecule Drug Design
• Recombinant DNA Drugs
• How is it done? What are the goals?
• What as been done so far?
• Antibody based drugs
• Vaccine Development and Production
• What is in the future?
• What are the controversies and concerns?
Recombinant DNA
And New Drugs
What Are the
Implications
Of Gene Cloning
For Pharma Now?
Genetic Constructs Now Make Proteins
That Are Pharmaceuticals
Promoter
Coding Sequence
Terminator
Your favorite gene
Controlled expression
“making protein”
Insulin
Enbrel
Herceptin
Stop transcription
Message stability
DNA Technology
and Pharmaceuticals
Allows precise treatments for:
Cancers
Cardiovascular diseases
Inflammation
Behavior
Obesity
Depression
Schizophrenia
Cracking the Genetic Code
(Nirumberg and Mathei 1962)
The information
is the same in all
living organisms
This fact is exploited for
DNA based drugs
Biomanufacturing Process for
Recombinant Proteins
Biomanufacturing
Vaccines and Antibodies
How Are They Made?
Controversies: Vaccines and Human Health: Myth Understood
Vaccines DO NOT Cause Autism
Why ALL Children Should be Vaccinated?
Can a Vaccine Compromise Your Immune System?
How Does Genomics Influence Vaccine Development?
Are we Ready for Contagion?
Why Are There Broad Public Misgivings
About Vaccination?
The Immune System:
What happens when humans are invaded
The Immune System:
Antibodies are proteins encoded by genes
Immunotherapy uses antibodies as drugs
Antibody Based Drugs
Promoter
Coding Sequence
Terminator
Coding sequence for an antibody
fragment to a specific antigen
Stop transcription
Controlled expression
Message stability
“making protein”
Monoclonal antibody therapy is the
use of monoclonal antibodies (or mAb)
to specifically bind to target cells or
proteins
Antibodies
versatile protein molecules capable of recognizing
foreign proteins
Fv
Antigen
Binding
Site
VH
CH1
Antigen/pathogen specificity
VL
CH2
Fc
Ancillary
functions
CH3
Effector functions:
• Neutralization/blocking effects
• Complement fixation
• Agglutination
The B cell receptor
Ag
Ag
b a
a
b
Production of Hybridomas
and Monoclonal Antibodies
Monoclonal
antibodies are a
homogeneous
population of
antibodies that are
specific to their
antigen
Western Blot Analysis
Home Pregnancy Test
Detection of specific proteins
proteins are separated
by gel electrophoresis
separated proteins are
transferred to a filter
antibodies recognize
proteins and are detected
with specific dyes
Applications of monoclonal antibodies
ELISA
Enzyme Linked
Immunosorbent Assay
Home Pregnancy Test an antibody test specific to
human chorionic
gonadotropin HCG is
captured and detected with
a color enzyme developed
product
Vaccines and Vaccination
Facts and Fictions
Vaccines and Antibodies
How Are They Made?
Controversies: Vaccines and Human Health: Myth Understood
Vaccines DO NOT Cause Autism
Why ALL Children Should be Vaccinated?
Can a Vaccine Compromise Your Immune System?
How Does Genomics Influence Vaccine Development?
Are we Ready for Contagion?
Why Are There Broad Public Misgivings
About Vaccination?
Vaccines and Vaccination
A foreign protein (antigen), an
attenuated, or killed virus will
stimulate the immune system to cells
with a memory for the antigen. The
immune system is activated when the
antigen is again encountered.
How Are Vaccines Made?
Vaccine Production
Methods
Example: Attenuated Vaccine
other Common methods
Flu vaccine timeline for production
Pandemic flu
Contagion
Other Vaccines
HIV Vaccine
HPV Vaccine
Bacterial Vaccines
Malaria Vaccine
How to Make a Vaccine - Six different ways
Similar-pathogen
vaccine: smallpox virus
Toxoid vaccine:
tetnus
Attenuated vaccine:
measles virus
Subunit vaccine:
hepatitis B
Killed vaccine: polio
virus
Naked DNA vaccine:
HIV virus
Attenuated vaccine: measles virus
Step 1 Use the tissue culture to grow new viruses
You are about to create a liveattenuated vaccine, which means that
you need to alter a pathogen—in this
case a measles virus—so that it will
still invade cells in the body and use
those cells to make many copies of
itself, just as would any other live
virus. The altered virus must be
similar enough to the original measles
virus to stimulate an immune
response, but not so similar that it
brings on the disease itself.
To create a new strain of the virus,
you’ll need to let it grow in a tissue
culture.
Attenuated vaccine: measles virus
Step 2 Fill the syringe with a strain of the virus that has
desirable characteristics
The tissue culture is an artificial
growth medium for the virus. You
will intentionally make the
environment of the culture different
than that of the natural human
environment. For this vaccine, you'll
keep the culture at a lower
temperature.
Attenuated vaccine: measles virus
Step 2 Fill the syringe with a strain of the virus that has
desirable characteristics - continued
Over time, the virus will evolve into
strains that grow better in the lower
temperature. Strains that grow
especially well in this cooler
environment are selected and allowed
to evolve into new strains. These
strains are more likely to have a
difficult time growing in the warmer
environment of the human body.
After many generations, a strain is
selected that grows slow enough in
humans to allow the immune system
to eliminate it before it spreads.
Attenuated vaccine: measles virus
Step 3 Completed vaccine ready for use
You have just produced a liveattenuated measles vaccine.
Like the smallpox vaccine, the virus
within the vaccine will invade body
cells, multiply within the cells, then
spread to other body cells. The virus
used in the measles vaccine today
took almost ten years to create. The
starting stock for the virus originated
from a virus living in a child in 1954.
Live-attenuated vaccines are also used
to protect the body against mumps,
rubella, polio, and yellow fever.
How Are Vaccines Made?
To Make a Bacterial Vaccine, First Sequence a Genome
‘You cannot develop
vaccines against a
bacterial pathogen
without the
genome.’
Is this
Science Fiction?
This couldn’t
really happenRight???
How accurate
is this?
How worried
should I be?
Pandemic: A Worldwide Outbreak of Influenza
How Are Vaccines Made?
Flu Vaccine Production Timeline
1.
2.
3.
4.
5.
6.
7.
Surveillance (year round) ID emerging new pathogens (i.e. H5N1)
Strain selection January - March
Manufacturing and production January - July
Purification and testing July - October
Filing and packaging July - December
Shipping August - November
Vaccination October and beyond
Questions:
Is this fast enough?
Can it handle a fast emerging new pandemic?
Can we make enough?
Who should get it first?
Issues in Biotechnology
During your time in college you will have what
number of sexual partners?
(A)
(B)
(C)
(D)
(E)
0-5
5-10
10-25
25-50
over 50
35
30
25
20
15
10
5
0
1
2
3
4
5
Issues in Biotechnology
Girls go First
During your time in college you will have what
number of sexual partners?
(A)
(B)
(C)
(D)
(E)
0-5
5-10
10-25
25-50
over 50
50
40
30
20
10
0
1
2
3
4
5
Issues in Biotechnology
Now the Boys
During your time in college you will have what
number of sexual partners?
(A)
(B)
(C)
(D)
(E)
0-5
5-10
10-25
25-50
over 50
30
25
20
15
10
5
0
1
2
3
4
5
Issues in Biotechnology
I support a mandatory HPV vaccine at age 12:
(A)
(B)
(C)
(D)
yes
no
undecided
there is still insufficient information
25
20
15
10
5
0
1
2
3
4
Recombinant
DNA Technology
and Pharmaceuticals
Antibody Targeting
Allows precise treatments for:
Cancers
Cardiovascular diseases
Inflammation
Behavior
Obesity
Depression
Schizophrenia
Antibody Based Drugs
Promoter
Coding Sequence
Terminator
Coding sequence for an antibody
fragment to a specific antigen
Stop transcription
Controlled expression
Message stability
“making protein”
Monoclonal antibody therapy is the
use of monoclonal antibodies (or mAb)
to specifically bind to target cells or
proteins
Antibody Based Drugs
What do all the names of these drugs have in common?
* Alemtuzumab
* Gemtuzumab ozogamicin
* Rituximab
They all end in -mab, shorthand for
* Trastuzumab
monoclonal antibody
* Ibritumomab tioxetan
They are all names of monoclonal antibody
based drugs targeted to cancer cells
Targeting proteins involved
in disease with precise
accuracy
Antibody Based Drugs
Drug delivery to precise
Targets
Lower side effects
Higher efficacy
Antibody Based Technologies
Antibody-nanoparticle drugs
magnetic
Antibodies for the future
radioactive
quantum dots
Antibody-nanoparticle devices
computers
oxygen delivery
communications
Rheumatoid Arthritis
Rheumatoid Arthritis is
an inflammatory disease.
It is an example of an
AUTOIMMUNE
DISEASE because the
victim’s own immune
system attacks a protein
of its own.
Rheumatoid Arthritis
Autoimmune diseases arise from an
inappropriate immune response of the body
against substances and tissues normally
present in the body. The immune system of
the patient ‘mistakes’ some protein as a
pathogen and attacks its own cells. This
may be restricted to certain organs (e.g. in
autoimmune thyroiditis) or involve a
particular tissue in different places (e.g.
Goodpasture’s disease which may affect the
basement membrane in both the lung and
the kidney). The treatment of autoimmune
diseases is typically with
immunosuppression—medication which
decreases the immune response.
Rheumatoid Arthritis is an Overreaction
of the Immune System
Immuno-Suppressant Treatments
In addition to common anti-inflammatory drugs such
as aspirin, which reduce pain, immuno-supressant
drugs often induce remission
1.
2.
3.
4.
5.
Gold Salts
Anti-malerials
Methotrexate (anti-metabolite)
Corticosteroids
Herbal treatments
Overstimulation of the Immune System
causes Inflammatory Disease
Macrophages, white blood cells that engulf foreign invaders, play
an important role in the body’s defense by making TNF alpha
and other “cytokines” which cause the inflammation
Chronic Activation of the “Acute” response to infection is
damaging to the body and must be halted
Painful
RECEPTORS are proteins on the surfaces of cells
that enable them to recognize each other
In 1881, Surgeon William Coley noted
that bacterial infections in cancer patients
sometimes caused tumors to become
“necrotic” or atrophy

In 1975 Anthony Cerami showed that
bacteria induced release of a “wasting”
factor in an infected host, a small protein
that also had anti-tumor activity

Issues in Biotechnology
Specialized proteins embedded in cell
membranes which receive and transmit
chemical messages are often desirable drug
targets and are referred to as:
(A)
(B)
(C)
(D)
(E)
random walkers
receptors
transgressors
retractors
diseases
Tumor Necrosis Factor - TNF α

In 1881, Surgeon William Coley noted that
bacterial infections in cancer patients
sometimes caused tumors to become “necrotic”
or atrophy

In 1975 Anthony Cerami showed that bacteria
induced release of a “wasting” factor in an
infected host, a small protein that also had antitumor activity

A role in Rheumatoid Arthritis and
inflammation (and other diseases)
ENBREL as an antibody mimics
the soluble TNF Receptor
Antigen
B-cell Receptors
Antibody
• By preventing the tumor
necrosis factor from
causing damage to the
tissue, the progression of
Rheumatoid arthritis is
slowed down
• Pain secondary to the
inflammatory process is
reduced also
Selected Recombinant Products For
Medical Problems Affecting Large
Patient Populations
Hemophilia
Benefix recombinant (FIX)
Wyeth
Rheumatoid Arthritis
Enbrel (etanercept)
Amgen
Kineret (anakinra)
Amgen
Remicade (infliximab)
Centocor
Issues in Biotechnology
Recombinate DNA technology has been able to
make what class of compounds as a new class
of effective drugs?
(A)
(B)
(C)
(D)
(E)
muscle fibers
antibodies and vaccines
lipids
Cox 2 inhibitors
stem cells
12
10
8
6
4
2
0
1
2
3
4
5
Selected Recombinant Products For
Medical Problems Affecting Large
Patient Populations
Hepatitis B
Engerix-B (recombinant hepatitis B vaccine)
GlaxoSmithKline
Intron A (interferon-a 2b)
Schering Corp
Recombivax-HB (recombinant hepatitis B vaccine)
Merck & Co., Inc
Acute Myocardial Infarction (heart attack)
Retavase (reteplase)
Centocor
TNKase (tenecteplase)
Genentech, Inc
The Immune System:
Antibodies are used as drugs that are
specific to their targets
Issues in Biotechnology
Rheumatoid arthritis is:
(A) an autoimmune disease
(B) totally eradicated
(C) curable with the correct diet
(D) has been most effectively treated with
homeopathic remedies
(E) a consequence of poor health habits
Viruses Used as Drug Delivery
Devices
Engineered to not be
pathogenic
Exquisite cell or target
specificity
(e.g. HIV specifically
targets T4
Lymphocytes)
Suicide delivery agents
Viruses Used as Gene Delivery
Devices
An approach to AIDS
Treatment?
(e.g. HIV specifically
targets T4
Lymphocytes)
Suicide delivery agents
Viruses for Gene Therapy
An approach to Gene
Therapy?
Cell specific gene delivery
Replacement of
defective genes
Cancer treatments
Addition of new genes
Genetic Surgery
8. Antibiotic resistant tuberculosis is on the rise world-wide.
Which approach to research treatment development would not
be the best choice?
(A) develop a vaccine using recombinant DNA technologies
(B) develop RNAi methods to target the tuberculosis bacteria
(C) develop cheaper methods to make the antibiotic
(D) develop early detection methods based on PCR
(E) sequence the tuberculosis genome to look for new drug
targets
9. Rheumatoid arthritis is:
(A) an autoimmune disease
(B) totally eradicated
(C) curable with the correct diet and vitamins
(D) has been most effectively treated with homeopathic
remedies
(E) best treated early with surgery
10. Specialized proteins embedded in cell membranes which
receive and transmit chemical messages are often desirable
drug targets and are referred to as:
(A)
(B)
(C)
(D)
(E)
random walkers
receptors
transgressors
retractors
transducers
11. Enzymes are:
(A)
(B)
(C)
(D)
(E)
only used in commercial detergents
genes involved with biochemical pathways
made primarily of lipid
not involved with energy production
usually proteins that catalyze reactions in cells
12. Recombinant DNA technology has been able to make what
class of compounds as a new class of effective drugs?
(A)
(B)
(C)
(D)
(E)
muscle fibers
antibodies
lipids
Cox 2 inhibitors
homeopathic treatments
13. What are the implications of gene cloning for the
pharmaceutical industry?
(A) technically a good idea but all candidates have failed in
Phase III trials
(B) it might work but it will never gain public acceptance
(C) drugs based on antibodies are now on the market made
using this technology
(D) technically a good idea but has yet to be proven
(E) none, it’s the materials of science fiction and Hollywood
movies
14. The ability to replace defective genes in a patient, as a sort
of genetic surgery, has not yet been effectively achieved and is
called:
(A)
(B)
(C)
(D)
(E)
gene therapy
chiral chemistry
combinatorial chemistry
recombinate drug technology
alternative therapy
15. HPV stands for:
(A)
(B)
(C)
(D)
(E)
High Purity Vaccine
Hallmark Pneumonia Vaccine
Henrietta’s Park Virus
Human Papilloma Virus
Human Pancreatic Virus
16. An influenza pandemic is a global outbreak of disease that
occurs when a new influenza A virus appears or “emerges” in
the human population, causes serious illness, and then spreads
easily from person to person worldwide. Such a pandemic:
(A) has only occurred once in recorded human history
(B) is only the material of Hollywood movies such as
“Contagion”
(C) is only a matter of time before another occurrence
(D) is totally preventable
(E) proves that the theory of evolution is incorrect since
viruses cannot evolve