biotechnology
TRANSCRIPT
An Introduction toBiotechnology
Amgen is a leading human therapeutics company in the biotechnology industry. Since our founding in
1980, we have focused on accomplishing our mission to serve patients by discovering, developing
and delivering innovative medicines to treat grievous illnesses. By pioneering the development of
novel products based on advances in cellular and molecular biology, Amgen’s therapeutics have
changed the practice of medicine and helped millions of people around the world to fight cancer,
kidney disease, rheumatoid arthritis and other serious illnesses.
Pioneering science delivers vital medicines
Chapter One: Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Chapter Two: The Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Chapter Three: How Biology Drives Biotechnology . . . . . . . . . . 9
Chapter Four: The Technology . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Chapter Five: Drug Discovery . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Chapter Six: Drug Development . . . . . . . . . . . . . . . . . . . . . . . . . 22
Chapter Seven: Scale-Up and Manufacturing . . . . . . . . . . . . . . 26
Chapter Eight: Biotechnology Medicines . . . . . . . . . . . . . . . . . . 29
Chapter Nine: Future of Biotechnology in Healthcare . . . . . . . . 31
Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Timeline of Medical Biotechnology . . . . . . . . . . . . . . . . . . . . . . 42
Table of Contents
In 1919, Karl Ereky, a Hungarian engineer, coined the term biotechnology* to describe the
interaction of biology and human technology. He envisioned a new era of technology based on
using biology to turn raw materials into socially useful products. Nearly a century later, Ereky’s
vision is being realized by thousands of companies and research institutions.
Chapter One:
Introduction
Introduction *Terms in boldface are defined in the glossary .2
Introduction
Modern biotechnology began in the 1970s
in Northern California and has since grown
into a worldwide industry . Amgen, founded
in 1980, was one of the first companies to
realize the new field’s promise by bringing
biotechnology medicines to patients .
Today, biotechnology industry sectors include
healthcare (biologics, devices, diagnostics),
agriculture (genetically modified organisms,
food safety), industry and environment
(biofuels, biomaterials, pollution) and biode-
fense (vaccines, biosensors). This booklet
focuses on healthcare .
Drugs
A drug is a therapeutic substance used to
prevent, manage or cure disease . In the
United States, the U .S . Food and Drug
Administration (FDA) must approve all drugs
before they are sold to the public . Most
countries follow global harmonized guide-
lines and have a regulatory agency similar
to the FDA that evaluates drug research
and approves drugs for marketing . The most
familiar type of drug is the synthesized drug,
such as aspirin . The pharmaceutical industry
traditionally manufactures synthesized drugs .
The biotechnology revolution brought about
a new class of drug: the biologic .
Biologics are therapies derived from living
organisms and include therapeutic proteins,
DNA vaccines, monoclonal antibodies and
peptibodies [a modality that combines the
active portion of a protein (peptide) with a
portion of the core structure of an antibody],
as well as new experimental modalities such
as gene therapy, stem cell therapy, antisense
nucleotides and RNA viruses .
Many biotechnology drugs are proteins.
Proteins, which are made from amino acids,
are the workhorses of the cell and perform
all functions within a cell . Because cells
produce proteins naturally, the biotechnology
industry utilizes cells, not chemicals, to
manufacture biologics .
BIoFaCT
To bring a new drug to market (from discovery through clinical trials and FDA approval) costs an estimated $1 billion and can take 10 to 15 years or longer .* Only one in 10 new drugs that makes it into human testing actually makes it to market . Given this high failure rate and the tremendous cost of bringing a new therapy to market, companies depend on successful drugs to produce enough revenue to compensate for both the R&D costs of the successful therapies and the expense of failed ones .
* Innovation .org . (February 2007) . Drug discovery and development: Understanding the R&D process [Brochure] . Washington, DC: Pharmaceutical Research and Manufacturers of America . 3
The Science
The biotechnology industry is based on living organisms. The cell is the basic unit of life. All living
organisms consist of one or more cells. Some organisms are unicellular, such as bacteria and
yeast. Others, such as humans, are multicellular, consisting of trillions of cells. All cells have
common processes they perform in order to survive. Biotechnology harnesses these processes
to make products to treat illness and improve health.
Chapter Two:
The Science
4
The Science
Cell Processes
Cells Replicate (mitosis): Prior to dividing,
a cell makes a copy of its DNA and other cell
parts . The cell divides and forms two identical
cells from the original single cell . These iden-
tical cells are referred to as daughter cells .
Cells Grow: After replicating, the daughter
cells grow to their intended size .
Cells Metabolize: Cell metabolism is the
process by which cells process nutrients and
maintain a living state . Cells break down large
molecules into smaller molecules to produce
energy and molecular building blocks, which
are used to create new cell structures and
control cell function .
Cells Respond to Stimuli: Unicellular and
multicellular organisms respond to internal
and external stimuli . For example, plants grow
toward a light source because light is needed
for photosynthesis and the production of
energy . Light and the ability to respond to its
presence are essential to a plant’s survival .
Cells can respond to a whole range of stimuli .
Cells Adapt: Organisms may thrive or die
based on their ability to adapt to adverse
environmental conditions such as changes
in temperature, solute concentration, oxygen
supply and the presence of hazardous agents .
Parts of an Animal Cell
The cell can be divided into three main sections:
the cell membrane, the cytoplasm (which
includes the organelles) and the nucleus.
Nucleus: The nucleus houses most of the
DNA and is the control center of the cell . It is
surrounded by a membrane that lets certain
molecules into and out of the nucleus to
keep the DNA safe . This DNA never leaves
the nucleus .
Cell Membrane: The cell membrane is the
border that surrounds a cell . It monitors what
goes into and out of the cell . Embedded in the
cell membrane are receptors. Receptors
cross the membranes and act as docking
stations for molecules . When a specific type
of molecule attaches to a receptor, a series
of chemical reactions (cell signaling) can occur
in the cell, creating a cellular response . If a
receptor is blocked, cell signaling is stopped
and no response occurs . This is how some
biologic therapies work: they attach to recep-
tors and interrupt cell signaling . Other biologics
work by mimicking the signaling molecule .
Organelles: There are many different types
of organelles within the cytoplasm of a cell, and
each performs a specific function . For exam-
ple, ribosomes make proteins . Mitochondria
make energy . The endoplasmic reticulum folds
and transports certain proteins . Golgi bodies
modify proteins and are involved in their
transportation around the cell . Vacuoles
store cellular waste products for disposal .
BIoFaCT
Ribosomes can assemble an average-size protein in one minute .
NucleusNucleusCell Membrane
Cell MembraneCytoplasm
Cytoplasm
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DNA
All cells contain deoxyribonucleic acid (DNA).
DNA is the blueprint for the construction
and operation of the cell . DNA is arranged
in large segments called chromosomes.
Within a chromosome are specific pieces
of DNA called genes. Genes vary in length
and contain the information a cell needs to
make proteins . Proteins control many aspects
of cellular function .
The information in DNA is stored as a code
made up of four basic “building blocks”
called nucleotides . DNA is very long, and
the order of the nucleotides determines the
information stored . Each nucleotide consists
of three components: a deoxyribose sugar, a
phosphate group and a base . There are four
different types of bases: adenine (A), thymine
(T), guanine (G) and cytosine (C) . The order of
the As, Ts, Gs and Cs in DNA gives meaning
to the cell, just as the order of letters in a word
gives meaning to that word and, by extension,
to a story in which the word appears . The
diversity of organisms is a result of the limit-
less combinations of bases—As, Ts, Gs and
Cs . Every organism contains DNA, but the
number and arrangement of bases are
different for every organism .
DNA is called a double helix because it is
posited to consist of two strands of nucle-
otides that bond together in a very specific
manner . The As bond with Ts, and the Cs
bond with Gs . The resulting A-T and C-G
combinations are called base pairs. In each
human cell, the length of DNA is equal to 3
billion base pairs . If flattened, a DNA segment
would look something like a ladder with two
side rails (phosphate and ribose groups)
and rungs (base pairs) between them . The
structure makes DNA very stable and able
to carry vast amounts of information .
Most cells within an organism contain the
exact same DNA, but not all genes within each
cell are active, or turned on . When a gene
gets turned on, the information encoded by
the gene is used to produce, or express, the
Determining the Structure of DNA In 1962, James Watson, Francis
Crick and Maurice Wilkins jointly
received the Nobel Prize in Physi-
ology or Medicine for discovering
the structure of deoxyribonucleic
acid (DNA). Because the Nobel
Prize is awarded only to the living,
Wilkins’s colleague Rosalind
Franklin, who died from cancer
at the age of 37, could not be
honored. But many attribute the
success of Watson and Crick’s
1953 discovery to Franklin, whose
X-ray crystallography images
of DNA helped them clarify the
structure of DNA.
BIoFaCT
Mitochondria are the only organelles in ani-mal cells to have their own DNA . Mutations in mitochondrial DNA can lead to illnesses such as Kearns-Sayre syndrome, which causes the loss of heart, eye and muscle movement functions .
The molecular structure of DNA—the double helix.
BIoFaCT
Humans have 23 pairs of chromosomes, for a total of 46—half inherited from the mother and half from the father . Chromo-somal abnormalities can involve numeric abnormalities, where there may be more or less than the normal 46 chromosomes, or structural abnormalities, where there may be duplications, translocations, deletions or even inversions of sections of certain chromosomes .
The Science
ChromosomeGene
DNA
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encoded protein . When a gene is not active,
or is turned off, it is not used to express
proteins . Depending on the cell’s function and
needs, genes are either turned on or off . Many
diseases are the results of genes improperly
turned on or off .
Mutations
Any change in the DNA sequence is called
a mutation . Environmental factors such as
exposure to radiation or chemical toxins can
cause mutations .
Mutations also can occur during the natural
process of DNA replication, when the cell is
responsible for copying 3 billion base pairs in
20 hours . Bases can be substituted, deleted
or repeated . Changes in the DNA sequence
may cause proteins to become dysfunctional
or may even, occasionally, improve function .
Genetic diversity results from an accumulation
of mutations over a long period of time, which
causes the differences among species .
Genomes
The term genome refers to the entire genetic
information in an organism . The human genome
is the entire DNA content found in a human;
the corn genome is the entire DNA content
found in corn; and so on . All genomes are
made up of the same bases: As, Ts, Gs and
Cs . The differences between genomes lie in
the number and sequence of base pairs and
the number and sequence of genes .
The number of base pairs does not correspond
to the number of genes; the two are indepen-
dent of each other . For example, the human
genome has 3 billion base pairs and approxi-
mately 20,000 to 25,000 genes . Only 3 percent
of the human genome codes for genes; 97
percent is termed noncoding DNA—in other
words, DNA that does not contain instructions
for creating proteins . Biologists have not yet
fully discovered the function of noncoding
DNA, but they suspect it may be involved in
the evolution of species or may have regulatory
functions within the cell .
Furthermore, the number of base pairs and
genes does not correspond to the intelligence
or physical capability of an organism . Compare
the human genome with the amoeba genome .
An amoeba, a unicellular organism, has the
largest-known genome—which may be as
large as 670 billion base pairs—as compared
to the human genome of 3 billion base pairs .
Yet a human is a more complex, intelligent
and physically capable organism .
Proteins
Proteins are made of long chains of amino
acids that fold into intricate and complex
3-D shapes . The order of amino acids is
determined by the DNA sequence in a gene .
There are 20 different amino acids . The se-
quence of amino acids determines the shape,
and therefore the function, of the protein .
The U.S. National Center for Biotechnology Information Established in 1988, the National
Center for Biotechnology Informa-
tion (NCBI) creates and maintains
extensive networks of biomedical
databases for storing and analyzing
genomes. The databases include
information about various genome
base pair sequences, genes and
proteins.
NCBI is a division of the U.S.
National Library of Medicine at
the National Institutes of Health.
See http://www.ncbi.nlm.nih.gov.
BIoFaCT
Variations in individual nucleotides occur within DNA at the rate of approximately one in every 1,300 base pairs in most organisms . In humans, the rate is one in every 1,200 base pairs . Most of these mutations, how-ever, do not adversely affect us; only a few are involved in the production of dysfunc-tional proteins or disease states .
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The Science 7
Protein synthesis is a complicated, multistep
process . Two of the steps involved with making
a protein are transcription and translation.
Transcription
During transcription, the original DNA code is
rewritten onto a molecule called messenger
RNA (mRNA). mRNA is also made up of a
sequence of nucleotides that differ slightly
from DNA nucleotides . RNA molecules are
very similar to DNA but have a ribose sugar,
are single-stranded and use U, or uracil,
instead of T, or thymine, as one of the four
possible bases .
Translation
During translation, the ribosomes assemble
individual amino acids into proteins . Ribo-
somes will bind to mRNA . Every three mRNA
nucleotides make up a codon—the code for
an amino acid . During translation, transfer
RNA (tRNA) reads mRNA and picks up the
corresponding amino acids . In this way, the
amino acids are linked together in a very
specific combination as dictated by the
sequence of nucleotides on the mRNA .
Short chains of amino acids are called
peptides. Long chains of amino acids are
called polypeptides . Polypeptides fold into
a functional protein .
Hundreds of different types of proteins per-
form specific jobs within and between cells .
Proteins called enzymes put molecules to-
gether and/or break molecules apart . Proteins
called signaling molecules allow cells to relay
messages to one another . Protein receptors
receive signals through a type of communica-
tion known as signal transduction or cell
signaling . Some proteins move substances
into and out of the cell . Structural proteins
give shape to cells and organisms . Proteins
are involved in cellular recognition and identify
different types of cells . Some proteins, such
as antibodies, are involved with defending an
organism against disease .
BIoFaCT
The highest-known number of genes in an organism is around 60,000—for the bacterium that causes trichomoniasis—which is almost three times as many as in the human genome .
The Science8
Biotechnology is based on biology, which is the study of life. The basic unit of life is the cell.
Biologists study the structure and functions of cells—what cells do and how they do it.
Biotechnologists use this information to develop products.
Biomedical researchers use their understanding of genes, proteins and cell parts to pinpoint the
differences between diseased and healthy cells. When researchers know how diseased cells are
altered and when they learn how to affect those alterations, they are better able to develop
innovative medical diagnostics, devices and therapies.
Chapter Three:
How Biology Drives Biotechnology
How Biology Drives Biotechnology 9
Understanding Disease Mechanisms
Early-stage drug research and development
(R&D) may begin with understanding the
underlying biology of a particular disease .
Biotechnology medicines are often created
specifically to address a particular disease
mechanism .
To design and develop new drugs, researchers
must understand the disease mechanisms
involved . Some initial questions researchers
ask to understand the underlying mechanisms
of a disease are: How does a person get
the disease? Which cells become diseased?
Is this disease caused by genetics, and if
so, what genes are turned on or turned off
in the diseased cells? What proteins are
produced—or not produced—in diseased
cells as compared to healthy cells? If the
disease is caused by a pathogen, what is
the interaction between the pathogen and
the person?
Studying disease mechanisms provides
researchers with information that can lead
them to identify targets for the early stages
of the drug discovery process . An under-
standing of fundamental biology may lead
to effective therapies for patients .
Take treatments available for autoimmune
disorders, for example . Autoimmune disorders
occur when a person’s immune system over-
reacts and attacks proteins, cells and tissues
in the body, often leading to inflammation .
Biologists have learned that tumor necrosis
factor (TNF) plays a major role in regulating
inflammation . Researchers know that too
much TNF is produced in autoimmune disor-
ders such as rheumatoid arthritis, psoriasis,
psoriatic arthritis, juvenile idiopathic arthritis
and ankylosing spondylitis . When too much
TNF is produced, excessive inflammation
occurs—and that can be damaging to joints,
skin and other parts of the body . Biotechnology
companies have worked to develop medicines
that inhibit the activity of TNF .
Models for Studying Disease
Researchers often take several different
approaches to creating models for study-
ing a particular disease . One approach is
to obtain samples of diseased cells and
healthy cells and grow them using a method
called cell culture. This calls for cells to be
incubated and fed with specialized growth
media. In culture, the cells do what cells
do—divide and express genes to produce
proteins . By studying how cellular processes
differ between healthy and diseased cells,
researchers hope to come to understand the
mechanism of disease .
Another approach involves studying shared
or similar genes and protein equivalents
in other species . Since all organisms are
made of cells and all cells perform many
similar functions, genes and proteins found in
humans are also found in other organisms .
The functions of human genes have been
revealed by studying parallel genes in
BIoFaCT
The process of analyzing and interpreting biologic scientific data is called computational biology and involves computer science, applied mathematics and statistics .
BIoFaCT
The biotechnology industry is one of the most R&D-intensive industries in the world . The United States is recognized as leading the world in biotechnology R&D .
Autoimmune Diseases Rheumatoid arthritis is a chronic
autoimmune disease that causes
inflammation and tissue damage
in joints and tendons. It can be a
disabling and painful condition
and can lead to substantial loss
of functionality and mobility.
Psoriasis is a noncontagious
disorder that causes red scaly
patches to appear on the skin.
These patches are areas of
inflammation and excessive skin
production.
Psoriatic arthritis is a type of
inflammatory arthritis that affects
up to 30 percent of people suffering
from psoriasis.
Juvenile idiopathic arthritis is
the most common form of chronic
inflammatory arthritis in children.
Ankylosing spondylitis is a
chronic, painful, degenerative
inflammatory arthritis primarily
affecting the spine and causing
eventual fusion of the vertebrae.
Lupus is a chronic inflammatory
disease that most commonly
affects women of childbearing
age. Lupus can affect the kidneys,
joints, blood and skin, among
other organs. Its symptoms can
include rash, fever, aches, anemia
and hair loss.
How Biology Drives Biotechnology10
nonhumans . This approach has added to
our understanding of how specific genes
and proteins direct the functioning of human
cells—both healthy and diseased .
Bioinformatics
Bioinformatics combines biology, computer
science and information technology into one
discipline . The goal of bioinformatics is to
capture, organize and index scientific infor-
mation so researchers can better understand
biology . The challenge for computer pro-
grammers is to design databases that allow
researchers to easily access existing data as
well as submit new data .
The scientific community generates volumes
of biological data daily . Biotechnology
companies use this information to form a
comprehensive picture of normal cell activity
so researchers can better study diseased
cells . This leads to the development of
diagnostic tools, therapies and preventive
medicines . Because of technological ad-
vances in biotechnology, bioinformatics has
evolved to focus on nucleotide sequences,
genes and amino acid sequences .
Biomarkers
Biomarkers are substances that can be
measured and evaluated to indicate (or serve
as markers of) normal biologic processes,
disease processes or biologic responses to
therapeutic treatment and disease intervention .
Historically, biomarkers were physiological
indicators—such as blood pressure or heart
rate . Today, disease may also be detected
using molecular biomarkers such as prostate-
specific antigen, a protein that if elevated,
may indicate prostate cancer .
Once a biomarker is validated (meaning all or
most patients with the disease test positive
for the specific biomarker), it can be used to
diagnose disease risk or presence of disease
or to help doctors determine patient treatment .
Biomarkers also can be used to predict
prognosis—for example, how a disease is
likely to progress if left untreated . Identifying
biomarkers that indicate specific disease is a
crucial step in the R&D process for developing
new biotechnology diagnostic tools .
BIoFaCT
Comparative genomics is the study of genome structure and function among different species . Researchers have obtained complete genomic sequences for the bacterium Escherichia coli (E. coli), the yeast Saccharomyces cerevisiae, the roundworm Caenorhabditis elegans, the fruit fly Drosophila melanogaster, the laboratory mouse and many other organisms for use in comparative genomic studies with the human genome .
KRAS: A Biomarker Breakthrough A biomarker can now help
physicians determine which
patients will not respond to a
certain class of anticancer agents
used to treat metastatic colorectal
cancer (mCRC). Amgen scientists
were the first to show in a phase
3 clinical trial that patients with
mCRC carrying specific cancer-
promoting mutations in a gene
known as KRAS are unlikely to
benefit from a drug designed to
bind to epidermal growth factor receptor (EGFR).
By testing for these mutations,
physicians can now eliminate
anti-EGFR antibodies as a treat-
ment option for patients with
mutated KRAS tumors.
KRAS research is regarded inter-
nationally as an important step
toward personalized medicine
for cancer treatment.
BIoFaCT
Genetic biomarkers are used in developing diagnostic tests for detecting certain genetic diseases . These tests look for DNA fragments identified as causing disease or associated with susceptibility to a disease .
How Biology Drives Biotechnology 11
Researchers study modifications and changes
that occur during cellular processes—such
as protein synthesis—in various disease
states . This can lead to the identification of
biomarkers that signal a change in disease
progression and can be used as a key devel-
opment in drug discovery and testing .
Biomarkers are also used in drug discovery to
determine whether a drug is effective in animal
models and at what doses effectiveness is
reached . Finally, biomarkers can be used in
disease management to determine whether a
drug is having the desired effect and whether
the correct dose is being used . This is very
important because response to a drug can
vary between patients: some require higher
or lower doses for the the drug to be effective .
Proteomics
The entire set of proteins produced by an
organism is called its proteome . Proteomics
is the study of protein structure and function .
Proteins control all aspects of cellular function .
Proteins produced in any specific cell of an
organism can vary with a number of factors
such as time, hormonal change and stress .
Proteomics researchers identify all proteins
involved with protein synthesis and protein
folding . Proteins need to fold correctly into
their 3-D shape in order to correctly function .
Even small structural defects during the folding
process can lead to a number of protein diseases .
Understanding protein structure, function
and interaction within and between cells is
crucial for drug discovery . Proteins are common
drug targets because they are responsible for
most normal cell functions—and malfunctions,
as in the case of disease . A drug target is
the molecule the drug interacts with to bring
about the desired change . This is sometimes
called hitting the target .
Cancer: From Biology to Treatment
Cancer research is a field that has been
at the forefront of utilizing bioinformatics,
biomarkers and protein studies to develop
new therapies that target specific cellular
processes . Cancer biology is a specific field
that explores all aspects of the cancer cell .
Researchers follow cancer pathways and
determine the molecular basis of cancer as
they develop diagnostics and treatments .
Cancer starts with changes in one cell or a
small group of cells that reproduce uncontrol-
lably . In healthy cells, cell division and growth
are tightly regulated . However, cancer cells
keep dividing—without normal checks and
balances—creating greater opportunity for
mutations . A mutation may result from the
environment or could have occurred during
DNA replication . As the cancer cells replicate
and grow, a mass (tumor) is formed . Cells
from the tumor can break away (metasta-
size) and spread through the bloodstream or
lymphatic system to other parts of the body,
creating new tumors .
Traditionally, cancer has been treated with
surgery, radiation and chemotherapy . The bio-
technology industry has contributed significant
advances in cancer treatment by developing
hormone therapies, biologics and targeted
therapies such as monoclonal antibodies .
Types of Cancer Cancer is a general term used
to describe diseases that occur
due to abnormal, uncontrolled
and rapid cell growth. There are
many types of cancer (carcinoma,
lymphoma, leukemia, sarcoma,
melanoma, etc.) that affect many
types of tissue (prostate, skin,
breast, brain, ovaries, lungs, etc.).
Carcinoma is cancer that begins
in the skin or in tissues that line
or cover internal organs. At least
80 percent of all cancers are
carcinomas.
Lymphoma is a cancer of the
lymphatic system—a network
of thin vessels and nodes
throughout the body that helps
to fight infection. Lymphoma
involves lymphocytes, a type
of white blood cell found in the
immune system.
Leukemia is cancer that starts in
blood-forming tissue—such as
bone marrow—and that causes
large numbers of abnormal blood
cells to be produced and to enter
the blood.
Sarcoma is cancer that begins in
bone, cartilage, fat, muscle, blood
vessels or other connective and
supportive tissue.
Melanoma is a cancerous
(malignant) tumor that begins
in the cells that produce skin
coloring (melanocytes). Melanoma
is almost always curable in its
early stages but can be lethal
in later stages.
BIoFaCT
Biotechnology has created more than 200 new biotherapeutics and vaccines, including products to treat cancer, diabetes, HIV/AIDS and autoimmune disorders . The majority of these products are therapeutic proteins .
How Biology Drives Biotechnology12
Biotechnology scientists depend on a wide variety of constantly evolving laboratory techniques and
tools. This section focuses on some of these platform technologies. To understand the industry, it’s
helpful to have some basic knowledge of what goes on in the lab.
Chapter Four:
The Technology
The Technology 13
Restriction Enzymes
Biotechnology employs a process called
genetic engineering, which combines DNA
sequences in order to produce recombinant
proteins as potential therapeutics . The process
utilizes restriction enzymes .
Scientists discovered restriction enzymes
(endonucleases) in bacteria in the 1970s .
They found that these enzymes cut up viral
DNA into small, nonfunctional pieces, thereby
protecting the bacterium from an invading virus .
There are hundreds of restriction enzymes—
each of them recognizing a specific sequence
of DNA called a restriction site . For example,
EcoR1, a restriction enzyme found in E. coli,
recognizes and cuts at the six-base sequence
GAATTC . HaeIII, a restriction enzyme found
in Haemophilus-aegyptius, recognizes and
cuts at the four base sequence GGCC .
All DNA, regardless of where it comes from,
is made up of the same four bases—As, Ts,
Gs and Cs . HaeIII reads any DNA segment
and cuts the DNA every time it encounters
the sequence GGCC . All restriction enzymes
are specific and reproducible, which are two
key characteristics that allow researchers to
utilize restriction enzymes to manipulate DNA .
The counterpart to cutting is called pasting .
DNA ligase is a protein (enzyme) that seals
two DNA segments together in a process
called ligation . The ability to cut and paste
DNA is the basis of genetic engineering .
Recombinant DNA
When segments of DNA are cut and pasted
together, the new DNA is called recombinant
DNA. Recombinant DNA can be inserted into
cells to produce cells with new characteristics .
This genetic altering can include a single-base
(letter) change or multiple gene changes .
Recombinant DNA can be introduced into
a host cell by a vector, which is used to
physically carry DNA into a host cell . A host
cell can be bacterial, yeast, plant, insect
or mammalian .
Common bacterial vectors include plasmids
and phages . A plasmid is a circular unit of
DNA that can be engineered to carry a gene
of interest . A phage is a genetically engineered
virus that injects DNA into bacteria . Cells that
contain recombinant DNA are referred to as
genetically modified, transgenic or transformed
cells . The process is called transformation.
+
BIoFaCT
Daniel Nathans, Werner Arber and Hamilton Smith received the 1978 Nobel Prize in Physiology or Medicine for their discovery of restriction endonucleases . Their discovery led to the development of recombinant DNA technology .
BIoFaCT
Scientists have identified more than 3,800 restriction enzymes, and more than 600 are commercially available for purchase from scientific supply companies .
Transformed CellsTransformation of bacterial cells
for the production of recombinant
proteins usually involves E. coli.
Transformation of animal cells,
called transfection, usually
involves a cell line derived from
Chinese hamster ovary (CHO)
cells. CHO cells were introduced
in the 1960s and remain the most
commonly used mammalian host
cells for industrial production of
recombinant protein therapeutics.
Restriction enzymes are proteins that function as molecular scissors and cut DNA.
The Technology14
The Technology
Recombinant Proteins
Recombinant DNA can be used to produce
recombinant proteins. The host cells use
the new DNA information and their own
cellular machinery to produce the protein
encoded by the recombinant DNA .
When recombinant proteins are produced for
use as human therapeutics, host cells must
be grown in large quantities so that enough
recombinant protein is produced to meet
demand . The recombinant protein is isolated,
purified and analyzed for activity and quality
before it goes to market .
Producing a protein with the proper order
of amino acids isn’t always the whole story .
Sometimes further processing is required to
yield an active or fully functioning protein .
Many human proteins are glycosylated,
meaning, they have a particular pattern of
sugar molecules linked to them . If a protein
is translated but not correctly glycosylated,
it may not function properly .
Adding a phosphate group—a process
known as phosphorylation—can act as an
on-switch, allowing the proteins to become
active proteins . Other biochemical functional
groups may be added to the protein to allow
it a larger range of function .
Recombinant proteins for therapeutic use
include vaccines, hormones, monoclonal
antibodies and hematopoietic growth factors
for the treatment of cancer, AIDS, aller-
gies, asthma and many other conditions .
The number of recombinant proteins has
increased greatly in recent years as the
technology used for their production and
purification has advanced .
Cell Culture
Cell culture is the technique of growing cells
in the laboratory under controlled conditions .
Growing large quantities of transformed cells
is a major step in the process of producing
recombinant protein products . Both
transformed bacterial cells and transformed
animal cells are used in this process .
Simple proteins can be produced using
recombinant DNA technology in bacterial cell
cultures . Typically, more-complex proteins
that are, for example, glycosylated are made
in animal cell cultures .
During cell culture, cells are grown in petri
dishes or flasks containing liquid media . Culture
media provides all the nutrients necessary
for cell growth . The cultures are grown in an
incubator that maintains the appropriate
temperature and environment, often requiring
certain gas mixtures of oxygen and carbon
dioxide . It is very important to maintain
specific conditions for cultures, because
variation in these conditions may affect
the proteins expressed and, therefore, the
product that is produced .
In the process of commercial production of
proteins, cell cultures must be scaled up to
produce enough protein to meet demand .
Since only a limited number of cells can be
grown in small petri dishes or flasks, the cell
culture can be transferred to large containers
called bioreactors, which are involved in the
manufacturing process .
BIoFaCT
The FDA approved human insulin in 1982—the first medicine made via recombinant DNA technology .
15
Research Tools
Biotechnology professionals depend on
leading-edge laboratory equipment . Here are
a few pieces used in genetic engineering:
Thermocycler
Polymerase chain reaction (PCR) is a
technique that replicates DNA in a machine
called a thermocycler. PCR is a series of
cycles that takes a small amount of original
DNA and exponentially copies, or amplifies, it .
Each three-step cycle doubles the amount of
DNA present . It’s like a photocopier for DNA .
A single piece of DNA can be turned into
millions of copies of the same DNA piece .
PCR enables researchers to make enough
DNA to work with in the lab . There are many
applications for PCR, including producing
enough of a DNA sequence or gene for use
in creating recombinant proteins .
Gel Electrophoresis
Gel electrophoresis is a technique commonly
used in the laboratory for analyzing DNA
fragments . Gel electrophoresis allows DNA
fragments to be separated within a gel . A gel
electrophoresis apparatus holds the gel and
allows electricity to run through it . Each DNA
fragment is negatively charged, causing it to
migrate toward the positive pole of the gel
apparatus . Larger fragments of DNA move
more slowly than smaller fragments because
they encounter resistance from the gel matrix .
There are different types of electrophoresis .
Each is based on the type of separating
material used, gels being one type, and the
types of molecules being separated . DNA,
RNA and proteins can all be separated using
electrophoresis in an appropriate apparatus .
Gel electrophoresis has many applications in
both clinical and research labs . One common
use is for verifying PCR products—that is,
checking to see whether the reaction generated
the correct DNA fragment .
DNA Microarrays
A DNA microarray (also called a gene chip)
is a small piece of glass or silicon divided
into thousands of sections in a grid pattern .
Each section has a single-stranded gene
fragment corresponding to either a healthy
or diseased gene .
DNA from an individual is separated into
single strands and tagged with a fluorescent
dye . The tagged DNA is washed over the
microarray . The individual’s DNA binds to
any complementary DNA sequences on the
slide, if they are present, to become double-
stranded DNA .
With the aid of a computer, the double-stranded,
fluorescently tagged DNA spots can be located
and measured . The individual’s DNA must
match the attached gene fragment exactly
to bind, thereby indicating the presence of
healthy or diseased DNA .
BIoFaCT
Kary Banks Mullis developed the polymerase chain reaction in 1983 and received the 1993 Nobel Prize in chemistry for the invention .
Gel electrophoresis.
The Technology
- +GelWell
16
Microarrays are powerful tools allowing
researchers to analyze thousands of genes
in one test . Microarrays are used for genetic
testing, for comparison of genetic information
from different individuals or species, and in
the discovery of potential drug targets .
Microarrays can also be used for researching
and identifying genes of interest to be used
with recombinant DNA technologies . Other
microarrays include protein, tissue, chemical
compound and antibody microarrays—all of
which allow researchers to analyze thousands
of data points at one time .
The Technology
Microarray.
BIoFaCT
Microarrays are capable of generating so much data that special microarray data-bases are needed for storage, searches, analysis and interpretation . There are both public and private microarray databases .
17
During drug discovery, scientists search for molecules—either chemical or biological agents—that
could alter a disease pathway. As part of the discovery process, they specifically look for ways to
change one or more molecular or cellular processes that occur in the affected cells of a diseased
tissue or organ.
Chapter Five:
Drug Discovery
Drug Discovery18
Drug Discovery
Chemical LibrariesIn the 1990s, chemists developed
huge libraries of chemical
compounds—thousands, even
millions, of chemicals with dif-
ferent structures used to screen
for new drugs. These libraries are
often proprietary and constructed
by a company explicitly to support
its drug discovery programs.
Initial screening of drug candi-
dates is relatively simple. Once
potential drug candidates or leads
are identified, more-complex
assays are used at subsequent
levels of screening. The sources
of the compounds found in the
drug candidates are often natural
products (from microbes, plants
and simple marine life) or chemi-
cal compounds synthesized by an
organic chemist.
Combinatorial chemistry increases
the potential of chemical libraries
by synthesizing larger, more
complex chemicals or chemically
related molecules from common
chemical structures. Combinatorial
chemistry for small-molecule
drugs includes the synthesis
of large organic molecules by
adding together smaller organic
molecules, often with improved
product results or lessened
product side effects.
Drug Discovery
Initiating Drug Discovery Research
An early step in the drug discovery process
is to identify an unmet medical need . What
is known about the disease? What are the
current treatment options, if any? Does the
company have the expertise, technology
and financial resources to solve the prob-
lem? Potential competitors and barriers, such
as regulatory constraints, are also taken into
consideration .
Target Discovery
After identifying an unmet medical need and
deciding whether it fits within the company’s
portfolio, scientists look very closely at the
biology behind the disease . Where can they
intervene, and what options do they have for
intervention? Since the human body is an
extremely complex system, scientists have
to carefully choose the target .
A target is a molecule that plays a critical role
in a disease . Scientists estimate that about
8,000 known therapeutic targets exist today .
Targets can be secreted factors, cell surface
receptors or signaling pathways within a cell .
The goal is to develop a drug that affects
a target in a way that interferes with the
disease process . It’s also very important to
ensure that the potential benefits of a drug
are appropriately weighed against any risks
such as possible side effects .
Different targets respond to different therapeutic
approaches . To select a target, scientists will
ask, “What are the differences between healthy
and diseased cells?”
Ultimately, disease processes take place at
the molecular level . There are various causes
of diseases . In inherited diseases, a difference
in the expression or in the sequence of genes
results in abnormal functioning of a person’s
cells . Sometimes this leads to a target being
present in excess; other times it could be
deficient or missing . So the scientists will
need to decide if the goal will be to block the
target or to enhance or replace it in order to
restore healthy function . For a disease caused
by an external pathogen, such as a virus or
bacterium, the pathogen produces molecules
that can damage the host organism’s cells .
Moreover, the pathogen will, itself, display
molecules in the infected individual that are
not present in a healthy person . The goal in
target discovery is to identify those differ-
ent molecules . This can be done using a
variety of technologies such as microarray
experiments, protein electrophoresis, mass
spectrometry (MS), DNA sequencing and
computerized imaging .
While this sounds straightforward, target
discovery is often difficult and may take years
to complete . Why? Cells and cell-to-cell
interactions are very complex . There may be
one or more mechanisms of the disease and
many points in the mechanism at which to
intercede . Moreover, the difference between
healthy and diseased cells can be too minute
to easily detect, or a method able to detect
the difference may not yet have been invented .
BIoFaCT
According to the National Center for Health Statistics, the top five diseases causing death in the United States in 2005 were heart disease, cancer, stroke, chronic lower respiratory disease and diabetes .
BIoFaCT
Researching the genetic and molecular basis of a disease is called studying the mechanism of disease .
19
The complexity of the body’s response also
means scientists could see a difference in
the expression of hundreds of genes without
being able to determine which ones were
critical to the disease .
Target Validation
Once scientists identify potential targets, the
next step is to validate them . Target validation
has two components . The first is to show
that the target molecule actually plays a role
in the disease . The second is to confirm the
target is a candidate for therapeutic interven-
tion: Can a safe and effective drug be made
against the target? Scientists complete this
second component of target validation before
the drug enters human testing .
There are a number of ways to validate a
target, and the process must take into con-
sideration time, cost and technology . At the
simplest level, the concept of target valida-
tion is to use the target to create the disease
in a sample of healthy tissues and then block
the target to restore the healthy condition .
This is done in cell culture or animal models .
The trick is to select a model that is repre-
sentative and will work . Sometimes people
who are born without certain functional
molecules express a specific disease type .
Studying biological samples taken from such
human subjects provides another means of
validating a target .
Examples of target molecules include recep-
tors, enzymes, ion channels, growth factors,
cytokines and DNA binding proteins . The
common thread among these targets is that
they are often involved in signal transduction
processes in and among cells . Signal trans-
duction pathways control cellular processes
such as division, differentiation, protein synthe-
sis and programmed cell death (apoptosis).
Initial studies are often done in cell culture . If
cell culture studies are positive, a next step
is to use an animal model .
Sometimes a suitable animal model has to be
created to validate a target . Sometimes the
target doesn’t exist in an animal model or may
not mimic the human disease state . Some-
times the drug candidate is so specific to
humans, it won’t recognize the animal model’s
target or the animal will mount an immune
response that blocks any therapeutic effect .
For example, Alzheimer’s disease occurs only
in humans, and only recently have mouse
models been developed to mimic the disease .
Scientists also look at what other effects the
drug candidate may have within preclinical
(both cell culture and animal) models . Some-
times the target is expressed on other cells or
tissues besides those directly involved in the
disease . What happens to those cells and
tissues in the presence of a drug candidate?
Does a drug candidate adversely affect other
cells or tissues? Does it raise an immune
response; stimulate other, similar targets; or
otherwise present any concerns about toxicity?
Preclinical work helps support later human
trials that may occur if the drug candidate
continues to show promise . Even if the drug
gets marketing approval after successfully
completing the necessary phases of human
Drug Discovery
Cell Receptors and Ion ChannelsThe most common drug targets
are cell receptors—proteins on
or inside a cell to which a specific
signaling molecule can attach.
These signaling molecules can be
hormones, neurotransmitters, pharmaceutical drugs, toxins or
even infectious agents. When
signaling molecules attach to
the receptor, a physical change
occurs that initiates a specific
cellular response.
Other common drug targets are
ion channels, proteins that form
pores in the membranes that
surround cells, and enzymes—
proteins that increase the rate of
specific chemical reactions.
BIoFaCT
Recently, scientists have begun using computer simulation to model drug-target interactions to guide drug discovery .
20
Drug Discovery
Choosing the Right Tool for the TargetDesigning a targeting strategy
usually comes down to a choice
between a small-molecule drug
and a biologic (most often a
recombinant protein or antibody).
Each has its particular advantages
and disadvantages.
Small molecules can usually cross
cell membranes and enter cells,
allowing them to be used for
targets inside cells. Biologics usu-
ally cannot cross cell membranes,
restricting their use to targets on
the surface of, or outside, cells.
Small molecules have good
specificity for their targets, but
recombinant antibodies generally
have extremely high specificity,
meaning, fewer adverse reactions
for the patient. Small molecules
have variable half-lives, which is a
measurement of how long a drug
stays active in the bloodstream
or in its target tissues. Biologics
often have much longer half-lives,
partly because they are modeled
on real biological molecules. This
means patients don’t have to
take as many doses of a biologic,
which may result in better patient
adherence to therapy.
Biologics usually need to
be injected, whereas small
molecules can be taken orally.
Small molecules can often cross
the blood-brain barrier, but
biologics usually cannot, which to
date has limited their usefulness
for treating diseases of the brain
such as psychiatric disorders and
neurodegenerative diseases.
trials, safety surveillance will continue once the
drug has reached the larger patient population .
Scientists will continue to answer safety
questions throughout the life of a drug .
Screening
High-throughput screening is a process
that combines robotics and data processing
to rapidly identify the compounds, antibodies
or genes that modulate a particular biomo-
lecular pathway . Large batches of potential
drugs are tested for binding activity or bio-
logical activity against target molecules .
Once a candidate disease is identified, a
company’s research lab develops a testing
method (assay) to determine or measure
the pharmacological activity of hundreds to
hundreds of thousands of molecules .
The assay measures the estimated potential
of a molecule to block or stimulate a target .
What’s being measured could be as simple
as the ability of the drug candidate to kill
cancer cells in culture or as complex as mea-
suring its ability to inhibit an enzyme involved
in a disease . Generally, the more complex the
assay, the more relevant the information—
but the higher the cost of the assay and the
longer it usually takes to get data .
Of the molecules that score a hit—that is,
a positive result that appears to have a
therapeutic potential—some are identified
as lead molecules due to their more druglike
properties (solubility, permeability, stability,
etc .) . Once a drug candidate is identified,
scientists may attempt to optimize its ability
to fight disease by changing its molecular
structure through combinatorial chemistry
for small molecules or protein engineering
for large molecules .
Drug Design
The design approach to drug discovery starts
with scientists understanding the genetic and
molecular base of a disease and using that
information to select a specific therapeutic
target . Drugs are then designed to interact
with the target . Through rational drug design,
scientists seek to develop a drug that is highly
specific to a particular target in a disease in
hopes of achieving a better therapeutic out-
come with potentially fewer side effects .
Scientists can learn more about the structure
of the target by using imaging technology
such as X-ray crystallography . 3-D structural
information about a target enhances drug
design strategies .
Considerations in designing a therapeutic
agent depend on both the nature of the target
and the capabilities of the company . If the tar-
get is on the exterior surface of the cell mem-
brane or is secreted, protein therapeutics such
as monoclonal antibodies or peptides can be
used . If the target is on the interior of the cell,
only drugs that can cross the cell membrane,
such as small molecules, can be used .
When designing a drug candidate, scientists
must keep in mind the intended method of
drug delivery and determine whether the drug
will be a pill swallowed, a liquid injected, a
spray inhaled or something else .
Drug Discovery 21
After the lengthy process of drug discovery (identifying a target and validating a drug candidate),
the process of drug development is still far from complete. Drug development includes the safety,
efficacy, formulation and manufacture of the drug. Typically, safety testing begins with a series
of experiments called preclinical studies. If these studies predict the drug candidate to be safe,
testing begins in humans in a series of studies called clinical trials.
Chapter Six:
Drug Development
Drug Development22
Drug Development*For more information on Amgen’s commitment to ethical use of animals in research, visit www .amgen .com/science/ethical_research .html .
Preclinical Studies
Preclinical studies are tests that take place
in a scientifically controlled setting using cell
cultures and animals as models . The goal of
preclinical studies is to predict what the body
does to the drug candidate (pharmacokinetics),
what the drug candidate does to the body
(pharmacodynamics), and whether the
drug candidate may pose potential health
hazards or toxic side effects .
Pharmacokinetic testing provides data to
answer questions such as: How is the drug
absorbed and transported? Which cells
and organs are affected? What enzymes
in the body break down the drug, and how
fast does this occur? How is the drug or its
metabolites (breakdown products) eliminated
from the body? Pharmacodynamic studies
examine dose-response effects and often
monitor biochemical and physiological changes
(such as enzyme activities, heart rate, blood
pressure and body temperature) in the test
subject . Pharmacodynamic testing, which
shows what the body does in response to
the drug, is used to answer the question:
Is the drug harmful or toxic to cells or organ
systems? Toxicology studies address the
potential of the drug or its metabolites to kill
or damage cells and organs, cause cancer or
cause reproductive problems, including birth
defects or sterility . Pharmacokinetic and
pharmacodynamic studies are used together
to reach the goal of preclinical studies, which
then answer the question: Is the drug safe?
In the United States, preclinical studies must
be conducted under FDA guidelines known
as current Good Laboratory Practice . Many
other countries follow global harmonized
regulatory guidelines as well .
Information from these studies is vital . It allows
researchers to estimate a safe dosage level
for humans in phase 1 clinical trials . Although
drug companies are required to submit animal
model data to regulatory agencies as part
of the drug approval process, companies are
taking steps to reduce the number of animals
used in testing because of ethical concerns
and the cost associated with facilities .
In the United States, institutes that conduct
research involving animals and that receive
federal funding must have an Institutional
Animal Care and Use Committee (IACUC) .
This committee reviews research protocols
and evaluates the care laboratory animals
receive . The IACUC is responsible for making
sure labs comply with the Animal Welfare Act .
Animal models greatly enhance scientists’
ability to test the effectiveness and safety of
new drug candidates . In target validation,
researchers may use knock-out mice and/or
knock-in mice to validate a target . Knock-out
mice are genetically altered to remove mouse
versions of human disease genes . Human
disease genes can also be knocked in to
create mouse models with human diseases like
cancer, diabetes, Alzheimer’s and Parkinson’s .
Drug candidates are tested on these mice,
enabling researchers to check for adverse
side effects before giving the candidate drug
to humans .
Initially, many studies of drug safety and
toxicity are done using cell lines. Cell lines
are engineered to express genes that are
often responsible for adverse reactions . The
creation of cell line models has decreased
the number of animals needed for testing
(reducing cost and time) and helps accelerate
the drug development process .*
23
Drug Development
How Is Dosage Determined? There are two types of phase 1
dosage studies: SAD studies and
MAD studies.
SAD: Single-Ascending-Dose Studies A few volunteers are given a
small dose of the investigational
new drug and observed. If there
are no adverse reactions, another
group is given a slightly higher
dose. This is repeated as many
times as needed until volunteers
start to exhibit intolerable side ef-
fects. At these dosage levels, the
investigational new drug is said to
have reached maximum tolerated
dose (MTD).
MAD: Multiple-Ascending-Dose Studies The same volunteers receive
higher and higher doses of the
investigational new drug until
the dosage reaches a certain
level. Samples of body fluids are
collected with each increase in
dosage level to understand
how the body processes the
investigational new drug.
Phase 1 clinical trials are usually
conducted in an inpatient clinic
where full-time staff can observe
the study subjects.
If preclinical trials provide sufficient evidence
that a drug candidate is safe, companies
submit an Investigational New Drug (IND)
application to the FDA . After the FDA approves
the IND, companies can begin phased clinical
trials in humans .
Clinical Trials
Clinical trials are tests designed to determine
the safety, proper dosage, efficacy, adverse
reactions and long-term-use effects of a new
drug in human subjects . Clinical trials taking
place in humans are conducted under global
harmonized guidelines, such as the FDA’s
current Good Clinical Practice (cGCP),
which protects the rights and ensures the
safety of human test subjects and follows the
U .S . Code of Human Research Ethics .
Clinical trials are conducted in three successive
phases—1, 2 and 3—and test progressively
larger numbers of humans in each phase . Each
phase has a different purpose and helps re-
searchers answer different questions . If a phase
is successful, the drug candidate proceeds to
the next phase . If unsuccessful, clinical trials
are halted, the drug is suspended and the
sponsor company returns to the discovery
phase to look for another drug candidate .
Clinical trials are conducted at different testing
sites . It takes several years to complete all
three clinical trial phases .
Clinical trials are often managed by a contract
research organization (CRO), which is an
independent organization . The CRO is
responsible for all the data management and
communication between the sponsor com-
pany and physicians overseeing the clinical
trials . The CRO also ensures that the study
volunteers understand and accept the risks
involved in the clinical trials and that cGCP
guidelines are followed .
Phase 1
Phase 1 trials represent the first time an
investigational new drug is tested on humans .
The goal is to evaluate the drug’s safety, tol-
erability and safe dosage range . The testing
group is often small, ranging from 20 to 50
volunteers . These are usually healthy volun-
teers who do not have a disease . However,
sometimes patient volunteers will be ac-
cepted into a phase 1 clinical trial, particularly
when testing oncology therapeutics . Usually
these patients have been unsuccessful with
available treatments or have few treatment
options, or the drug’s potential side effects
are too risky to involve healthy subjects (such
as using some chemotherapeutic agents) .
Phase 2
The goal of phase 2 trials is to determine
the efficacy and safety of the investigational
new drug among a larger group of patient
volunteers—usually 100 to 300 people .
A patient volunteer is someone who has the
disease the drug is intended to treat . Some
companies divide phase 2 trials into phase
2A (to assess dosage) and phase 2B
(to assess efficacy) . Most investigational
new drugs fail during this stage because
of efficacy and/or safety issues .
BIoFaCT
A crucial component of initiating a clinical trial is recruiting study subjects who agree to participate and sign a document called informed consent . Potential subjects must be informed about all aspects of the study before they decide to participate . Participants can withdraw their informed consent at any time .
24
Drug Development
Phase 3
The goal of phase 3 trials is to confirm the
effectiveness of the investigational new
drug and compare it with placebos or
therapies already available on the market .
To do this, hundreds or thousands of patient
volunteers are tested . Phase 3 trials are the
most expensive and time-consuming, lasting
for a couple of years or longer to establish
long-term safety .
Once phase 3 is successfully complete, the
sponsor company files a new drug or biolog-
ics application with the country’s regulatory
agency . In the United States, the company
would file a New Drug Application (NDA) for a
small-molecule drug or a Biologic License
Application (BLA) for a large-molecule drug
with the FDA . If the governing regulatory au-
thority (the FDA in the United States or the
European Medicines Agency, known as the
EMEA, in Europe) approves the drug, the
sponsor company is permitted to market and
sell the product in the country or countries
regulated by that authority . The final manufac-
turing of the drug—or large-scale production—
must take place in a facility that meets the
country’s strict guidelines, such as the FDA’s
current Good Manufacturing Practice (cGMP),
to ensure safety and purity of the product .
Phase 4
Phase 4 trials occur after an approved drug is
on the market . A goal is to monitor the drug’s
safety and efficacy when utilized in a normal
medical setting in a population of patients
that could number in the millions . Sometimes
adverse reactions, which were not seen in
a comparatively small cohort of patients
(3,000 patient volunteers as compared to
millions), are discovered in larger and more
diverse populations . If an adverse reaction is
discovered, the drug may be withdrawn from
the market . Either the sponsor company can
voluntarily withdraw the drug or a regulatory
body can pull the drug from the marketplace .
After further testing, the drug may or may not
be reinstated .
The stages in product development, or
product pipeline, take, on average, 10 to
15 years to complete . Most investigational
drugs do not make it . Out of every 1,000
potential new drugs in discovery, only one
will make it to approval .
Study DesignsLate-stage trials often include
a double-blind randomized
controlled test. In this type of
study, neither the patient volun-
teer nor the researcher knows
which volunteer belongs to the
control group or the experimental
group. Each patient volunteer is
randomly placed into one of the
groups. A third party keeps this
documentation and releases it
only after the study is over.
BIoFaCT
One of the largest challenges associated with clinical trials is the shortage of study subjects .
25
The manufacturing of biologics is complex, since most are proteins—large molecules often variable
in structure and sensitive to environmental conditions. The manufacturing of biologics has become
a science that can be summarized in four key steps: producing the master cell line, growing cells
and producing protein, isolating and purifying protein from cells, and preparing the biologic for
patients. The whole process from creating the master cell bank to preparing the biologic for
patients can take years and cost hundreds of millions of dollars.
Chapter Seven:
Scale-Up and Manufacturing
Scale-Up and Manufacturing26
Cell BanksCell banks involve a two-tiered
frozen cell banking system: a
master cell bank (MCB) and a
working cell bank (WCB). Scien-
tists use one vial of cells from
the MCB to create the WCB. Once
established, the WCB is used to
produce batches of product in
the scale-up process. Working
from the same stock of cell line
reduces the chance of mutations
associated with serial cultures.
The MCB is a reserve of cells
that scientists use only when
absolutely necessary. To protect
the integrity of the cell lines,
companies store their cell banks
in two or more locations within
their facilities and in one location
off-site.
Growth vessels vary in size:• Flaskshold5mL
• Spinnerflasksorrollerbottles
hold 50 to 200 mL
• Benchtopbioreactorshold
5 to 20 liters
• Pilotscalebioreactorshold
50 to 200 liters
• Productionvesselshold
20,000 liters or more
Using R&D Specifications
During the R&D phase, researchers develop
the initial production methods on a small scale .
They also determine the drug’s final formula-
tion, or physical form, for clinical trials—for a
biotechnology medicine, usually an injection
or infusion . Using all of the R&D data from
these production steps, companies devise
large-scale production methods to produce
enough of the product for the intended market .
The scale-up and manufacturing process must
adhere to cGMP guidelines to ensure product
safety and purity .
Common Cell Lines
Many biotechnology products are proteins
that must be produced by cells grown in
culture . Chinese hamster ovary (CHO)
cells, nonsecreting (NS0) cells (pronounced
“NS zero”) and E. coli are cell lines used for
production of biotherapeutics, especially
monoclonal antibodies .
There are a number of reasons to use these cells .
Both CHO and NS0 cells synthesize proteins
much like human cells do . Both are immortal
cell lines, meaning, they should be able to
grow and produce product forever . Researchers
are well versed in their optimal culture conditions .
Both cell lines have generally regarded as
safe (GRAS) status for therapeutic protein
production . NS0 cells have the additional
advantage of being programmed to produce
antibodies, but they do not make or secrete
any of their own antibody protein .
Other cell lines can be used and may be
more suitable . The selection of the cell line
depends on the expertise of the company,
the properties of the cell and the regulatory
requirements .
Scale-Up Process
The scale-up of a cell culture process can be
very difficult and time-consuming, taking as
long as several months before researchers
can obtain a product . The entire process of
producing a biotech product from start to
finish is often called a campaign and is usually
divided into two main parts: upstream and
downstream. Upstream processes involve
production of the protein product, most often
by using cells (microbial, insect or mammalian)
growing in culture . Downstream processes
include the recovery, purification, formulation
and packaging of the protein product .
Upstream Phase
Upstream processing begins with the cells
that scientists create or engineer to make
the protein product . Once the desired cell
line is made, it is cryopreserved: scientists
freeze a large number of vials of the cells to
create a cell bank. To begin a campaign,
scientists remove and thaw a vial of cells
from the cell bank and initiate a cell culture
in a flask containing a small volume of
growth media . The initial volume of media
can be as little as 5 mL . The media provides
the nutrients and the optimum environment
for cells to survive .
Scale-up is done by gradually transferring the
growing cells into successively larger growth
vessels containing larger media volumes .
The cells are constantly dividing as long as
the growth environment remains favorable .
Therefore, more and more cells are present
BIoFaCT
The American Type Culture Collection (ATCC) is a private, nonprofit resource dedicated to the collection, preservation and distribution of authentic cell lines and other biologic materials . (www .atcc .org)
Scale-Up and Manufacturing 27
with each step . The greater the number of
cells, the more protein product is generated .
Scale-Up Monitoring
The goal of the scale-up process is to grow
cells as quickly as possible and to produce
as much protein product as possible .
Using the same assays or testing methods
used in the initial R&D stages, scientists
measure cell viability and concentration,
product concentration and product activity
at each incremental scale-up stage for
monitoring purposes .
Lab technicians monitor and control the
physical environment in which cell cultures
grow . They do this manually in the initial
scale-up steps to optimize growth param-
eters such as temperature, pH, nutrient
concentration and oxygen level .
The monitoring process is automated once
the cell culture is large enough to be grown
in bioreactors .
It is crucial during the scale-up, fermentation
and manufacturing stages that technicians
monitor and test the cultures for contamination
by bacteria, yeast or other microorganisms .
Any contamination of a culture ruins the entire
batch of product and costs a company money
and time . Technicians follow very strict protocols
for maintaining aseptic conditions at all times
during the scale-up and manufacturing stages .
Quality Control and Quality Assurance
Quality control (QC) and quality assurance
(QA) departments are responsible for all of
the monitoring that is crucial to the success
of the scale-up and manufacturing stages of
product development . The QC department
assures product quality and testing during
the product development stages well before
the product is at the stage of marketing,
ensuring that the scale-up and manufacturing
processes meet certain standards . The QA
department is usually responsible for meeting
and reporting quality objectives .
Downstream Phase
In the downstream phase of manufacturing,
the protein product is isolated from the cells
that produced it . Proteins found inside the
cell (intracellular proteins) require special
protocols to extract them for purification .
Usually this involves bursting the cells open
to release the protein product, which then
has to be purified away from the other com-
ponents that were inside the cell . Proteins
found outside of the cell (extracellular
proteins) can be easier to isolate .
After harvesting the protein product, the
next step is clarification. This is where
scientists separate the protein from cellular
debris . Then they apply the protein solution
to a series of chromatography columns to
obtain a pure protein product . Purification of
protein mixtures by column chromatography
separates proteins based on physical and
chemical properties such as size, shape or
charge (+ or –) . Additional purification steps
remove any residual DNA and deactivate any
viral particles that may be present .
Researchers verify the isolation and
purification of the protein product through
confirmed testing protocols . The protein
product is then formulated according to
the R&D specifications and packaged for
use by physicians and patients .
Scale-Up and Manufacturing28
Biotechnology Medicines
The biotechnology industry uses advanced technologies to apply cellular and molecular biology
to create new, beneficial products. Medical biotechnology products are used to treat or prevent
diseases. These products include therapeutic proteins, monoclonal antibodies, vaccines, allergy
immunotherapy products, blood components and tissues and cells for transplantation.
Chapter Eight:
Biotechnology Medicines
29
Biotechnology Medicines
Therapeutic Proteins
Scientists use recombinant DNA technology
to make therapeutic proteins, often referred
to as biologics . Biologics are used in such
fields as oncology, rheumatology, immunology,
endocrinology and virology . Approximately
50 recombinant therapeutic proteins are
approved for clinical use and are currently
marketed, and hundreds more are undergoing
clinical trials . Some biologics have been in
use for more than 20 years and are considered
standard therapy .
Doctors have long used therapeutic proteins to
replace or supplement patients’ natural body
proteins—especially when natural protein
levels are decreased or lost due to disease .
Some recombinant proteins are versions of natural
body proteins, and other versions are not exact
versions but produce similar effects in the body .
Vaccines
Vaccines stimulate the immune system and
provide protection against particular diseases .
The first vaccines were made with inactivated
(killed) or weakened virus unable to reproduce
in the body but sufficient to provide immunity
upon future exposure to the live virus .
Vaccines are also created with recombinant
proteins . Scientists use genetic engineering
to create recombinant vaccines by inserting
genes for desired antigens into a vector . A
vaccine vector, or carrier, is a weakened virus
or bacterium into which harmless genetic ma-
terial from another disease-causing organism
can be inserted . Typically, the body recognizes
antigens as foreign, and white blood cells will
attack them . Recombinant vaccines, however,
do not cause disease but do have the antigen,
thus tricking the body into thinking it is being
attacked by a pathogenic virus . Recombinant
vaccines are safe and easily grown and stored .
Antibodies
A major area of biologics is the production
of humanized or fully human antibodies .
Antibodies can attach to antigens found on a
pathogen and flag the pathogen for destruction
by the immune system . Antibodies also can
attach to proteins on immune cells that are
involved in autoimmune responses in diseases
like rheumatoid arthritis and multiple sclerosis .
Humanized antibodies are engineered to be
mostly human to avoid problems with rejection .
Fully human antibodies are derived from human
cells or human antibody genes .
Peptibodies
Peptibodies are engineered therapeutic fusion
proteins with attributes of both peptides and
antibodies but are distinct from each, and bind
to human targets .
Diagnostics
In addition to recombinant proteins being used
as biologic drugs, scientists use recombinant
DNA technology to produce a number of
diagnostic tests for diseases, including tests
for hepatitis and AIDS . In fact, scientists
commonly use recombinant protein antigens
as diagnostic reagents in enzyme-linked
immunosorbent assays (ELISAs) for the
detection of infectious agents such as Severe
Acute Respiratory Syndrome (SARS) .
Monoclonal AntibodiesThough monoclonal antibody
technology was invented in the
mid-1970s, it took 20 years
before the technology showed
its true potential. The first
experimental monoclonal antibod-
ies developed in mouse models
were ineffective because the
human immune system rejected
mouse antibodies as foreign. The
subsequent development of first
humanized, then fully human,
antibodies has enabled the suc-
cessful use of this breakthrough
technology in fighting cancer and
other serious illnesses.
Biotechnology Medicines30
Future of Biotechnology in Healthcare
Biotechnology can offer patients more and better healthcare choices. New, innovative diagnostics
and therapies are changing how some human diseases are prevented and others are treated. This
monumental healthcare shift is in its early stages, with novel medicines, diagnostics and technologies
in development that hold great potential to improve patients’ lives.
Chapter Nine:
Future of Biotechnology in Healthcare
31
Future of Biotechnology in Healthcare
Personalized Medicine
Personalized medicine is the concept that
patients should be treated with therapies and
medicines based specifically on each patient’s
unique genetic makeup, for optimal results .
Currently, the practice of medicine is based
on standards of care that are determined by
averaging responses across large groups of
people . Personalized medicine is a new para-
digm that proposes to manage a patient’s
disease based on the individual patient’s spe-
cific characteristics, including age, gender,
height, weight, diet, genetics and environ-
ment . Genetic testing is beginning to allow
the development of genomic personalized
medicine—medical care based on a patient’s
genotype or gene expression profile .
Pharmacogenomics
A major movement in healthcare is phar-
macogenomics. Pharmacogenomics takes
advantage of the fact that individuals have
unique genomes representing their genetic
makeup . Each genome is likely to react differ-
ently to a particular drug and dose amount .
The challenge is to identify which drug and
which dose will work most optimally for
each person or for groups of individuals who
share similar genetics . By understanding a
patient’s genetic makeup, a physician can
better prescribe a drug and dose level that will
optimally work to combat a particular disease .
Advances in DNA technology are the keys to
both pharmacogenomics and personalized
medicine . These advances allow for testing
and identifying an individual’s unique genetic
makeup and then comparing those differenc-
es with the population at large . Knowledge of
the human genome, variations of the genome
among individuals and variations of the encod-
ed proteins produced enables researchers to
develop medicines that address the individual
needs of each patient . Pharmacogenomics
and personalized medicine promise to improve
clinical trials for new drugs, advance screening
technology for diseases and result in more-
effective individualized healthcare and advances
in preventive medicine .
Genetic Testing
The biotechnology industry has brought about
vast improvements in testing and diagnosis for
genetic diseases . The discovery of single-
nucleotide polymorphisms (SNPs)—single-
nucleotide changes in the DNA sequence—
was one of the major breakthroughs in genetic
testing . SNPs (pronounced “snips”) represent
one of the most common forms of genetic
variation among individuals . When a SNP
occurs in a gene sequence that encodes for
a specific protein, it may change that protein
and cause a disease or increase a patient’s
susceptibility to a disease . Utilizing technology
to detect SNPs allows for more-accurate
diagnosis of genetic diseases and therefore
facilitates treatment decisions . Genetic testing
provides patients with both an understanding
of possible risks for certain diseases and
possible opportunities for prevention .
BIoFaCT
Molecular diagnostic tests analyze DNA, RNA or protein molecules to identify a disease, determine its course, evaluate responses to therapy or predict individual predisposition to a disease .
BIoFaCT
Approximately 10 million SNPs have been identified in the human genome .
32
Gene Therapy
Gene therapy is an emerging area of applied
genetics that utilizes recombinant DNA tech-
niques . In this case, the recombinant DNA
molecules themselves are used for therapy .
Gene therapy involves inserting genes,
created by recombinant DNA technology,
into the cells and tissues of patients to treat
their diseases . Scientists are studying gene
therapies for a number of inherited human
diseases involving defective genes . The idea
is to replace them with new, functional genes .
Since the first clinical trial was initiated in
1990, gene therapy research has expanded
greatly, with an increasing number of human
trials . The field, still in experimental stages,
focuses its efforts on patients with severe
and life-threatening diseases who usually
have few treatment options or who have
failed all available therapies .
Stem Cells
Stem cells are unspecialized cells that can
renew themselves indefinitely to produce more
stem cells . They can mature and develop
specialized functions or differentiate under
specific growth conditions . Stem cells eventually
differentiate to form all of the different types of
cells that make up the body . The broad potential
of an undifferentiated stem cell to make a variety
of other cells is the focus of stem cell research .
Stem cell therapy, which is still in experimental
stages, involves growing stem cells in the lab
and guiding them toward a desired cell type by
adding different growth factors . The differenti-
ated cells are then surgically implanted . The
theory is that stem cells may then integrate
into the diseased tissue, replace diseased
cells and reverse the effects of the disease .
Cell therapies also could be developed in
which undifferentiated stem cells may be
implanted along with growth factors to guide
their differentiation in the patient’s body . The
aim is to replace the damaged cells with
healthy, disease-free cells—hence the term
regenerative medicine for this approach .
The hope is that stem cells, directed to dif-
ferentiate into specific cell types, could be a
renewable source of replacement cells and
tissues used to treat a wide range of diseases .
Nanotechnology
Nanotechnology deals with the manipulation
of molecules and structures on a nanometer
(one-billionth of a meter) or atomic scale .
Applying nanotechnology for the improvement
of human health is called nanomedicine.
Biotechnology nanomedicine harnesses living
organisms and/or their components on a very
small scale .
One example of nanomedicine is the experi-
mental use of nanoshells to selectively target
and destroy cancer cells at the cellular level .
Nanoshells are nanoscopic metallic lenses
that are selectively delivered to specific
organs or tumors through the bloodstream .
Nanoshells have the ability to capture infrared
light shown through the skin of a cancer patient
and convert it to heat, which kills only the
targeted cancer cells .
Nanoparticles called buckyballs—uniquely
shaped and constructed carbon molecules—
are also showing potential for drug delivery to
target molecules or cells . They may make it
possible to deliver drugs that do not dissolve
in water . Also, because of their small size,
they allow more of the drug to be delivered
per volume . Scientists are working on nano-
particles to unclog blocked arteries .
Future of Biotechnology in Healthcare 33
Future of Biotechnology in Healthcare
New Drug Delivery Systems
Biomedical researchers are studying new
ways of delivering drugs within the body that
could improve effectiveness . One example
is the development of microscopic particles
called microspheres that have tiny holes just
large enough to carry and deliver drugs to
their targets . They are made out of materi-
als that resemble naturally occurring fats in
cell membranes and are delivered as a mist
sprayed into the nose or mouth .
Microsphere therapies are currently available
for lung cancer and respiratory illnesses .
Current research is investigating the use of
microspheres to deliver anticancer drugs to
active tumors and for use with anesthetics
in pain management .
34
The practice of medicine has changed dramatically over the years through pioneering advances in
biotechnology research and innovation, and millions of patients around the globe continue to benefit
from the treatments developed by companies that are discovering, developing and delivering innovative
medicines to treat grievous illnesses. As companies continue to develop medicines that address
significant unmet needs, future innovations in biotechnology research will bring exciting new advances
to help millions more people worldwide.
Looking Ahead
Future of Biotechnology in Healthcare 35
Glossary
Amino Acids: The building blocks of proteins . The unique sequence of amino acids in a chain defines the character of a protein molecule .
Angiogenesis: The process by which the body forms and develops new blood vessels . An-giogenesis can be both beneficial and harmful; while it can be used to stimulate development in new blood vessels to fight clogged arteries, it also allows malignant tumors to increase in size . Angiogenesis is a key area of cancer research .
Antibody: A component of the body’s immune response . A Y-shaped protein, it is secreted in response to an antigenic stimulus . It neutralizes the antigen by binding to it .
Antigen: Any substance, almost always a protein, not normally present in the body that when introduced to the body stimulates a specific immune response and the production of antibodies .
Apoptosis: The process of programmed cell death that may occur in multicellular organisms . Programmed cell death involves a series of biochemical events leading to characteristic cell changes and death . Apoptosis is a key area of cancer research .
Aseptic: Describes a product or method free of microbiological organisms that would lead to contamination .
Assay: A test procedure whereby a property or concentration of a substance is measured .
Autoimmune Disorders: Diseases whereby an individual’s immune system mounts an attack on a portion of its own tissues . Tissues undergoing such an attack can be destroyed in the process .
Base Pairs: Two nucleotides on opposite complementary DNA or RNA strands that are connected via hydrogen bonds . In DNA, adenine (A) forms a base pair with thymine (T), as does guanine (G) with cytosine (C) . In RNA, thymine is replaced by uracil (U) .
Bioinformatics: The application of information technology to the field of molecular biology . Bioinformatics entails the creation and advance-ment of databases, algorithms, computational
and statistical techniques, and theory to solve formal and practical problems arising from the management and analysis of biological data .
Biologic: A product derived from a living organism (from animal products or other biological sources) that is used in the diagnosis, prevention or treatment of disease . Examples of biologics include recombinant proteins, allergy shots, vaccines and hematopoietic growth factors .
Biologic License Application: An application filed with the FDA seeking approval to market a novel biologic in the United States . The application contains a description of the trials and results, formulation, dosage, drug shelf life, manufacturing protocols, packaging information, etc .
Biomarker: A substance used as an indicator of a biologic state . It is a characteristic that is ob-jectively measured and evaluated as an indicator of normal biologic processes, pathogenic pro-cesses or pharmacologic response to a specific therapy . Biomarker identification and measure-ment are regarded as key developments for the future of disease treatment . Biomarkers are also used in drug discovery to determine whether a drug is effective in animal models and at what doses effectiveness is reached .
Biopharmaceutical: A synthetic drug produced utilizing certain biotechnology methods .
Bioreactor: A device or system for growing cells or tissues in the context of cell culture . The pro-cess of fermentation is performed in a bioreactor to grow large volumes of cells for producing specific proteins .
Biosensor: A device that combines a biological component with a physicochemical detector component to detect a pathogenic agent .
Biotechnology: Technology based on biology, especially when used in agriculture, food science and medicine . The United Nations Convention on Biological Diversity defines biotechnology as “any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use .”
Glossary
36
Glossary
Blood-Brain Barrier: A physiological mechanism that alters the permeability of brain capillaries so that some substances, such as certain drugs or toxins, are prevented from entering brain tissue, while other substances are allowed to enter freely .
Cell Bank: A facility where cell lines are kept frozen and stored for later use . Cell banks include master cell banks (MCBs) and working cell banks (WCBs) . MCBs house primary cell strains that are kept stored and not used for production purposes . WCBs house cells used in pharmaceutical production grown from those maintained in an MCB so that their stability and uniformity are well characterized .
Cell Culture Technology: The growing of cells outside of living organisms . With mammalian cell culture, it is sometimes possible to replace animal testing with cell testing when evaluating the safety and efficacy of medicines .
Cell Lines: Generations of cells grown from original primary cells . Primary cells are cultured directly from a living organism . With the exception of some derived from tumors, most primary cell cultures have limited life spans . After a certain number of population doublings, cells usually stop dividing, though they remain alive . An established or immortalized cell line has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification .
Cell Viability: Determining whether a cell popula-tion is living or dead . Testing for cell viability usu-ally involves looking at a sample cell population and staining the cells or applying chemicals .
Chemical Library (or Compound Library): A collection of stored chemicals that may be used in high-throughput screening for drug development . The larger the chemical library, the better the chance that high-throughput screen-ing will find a hit (a potential drug candidate) .
Chinese Hamster Ovary Cells (CHO cells): A cell line often used in biological and medical re-search, first introduced in the 1960s . CHO cells are used in studies of genetics, toxicity screen-ing, nutrition and gene expression, particularly expression of recombinant proteins . CHO cells are the most commonly used mammalian hosts for industrial production of protein therapeutics .
Chromatography: A process by which complex
mixtures of different molecules may be sepa-rated from each other . This is accomplished by subjecting the mixture to many repeated partitionings between a flowing phase and a stationary phase .
Chromosome: A threadlike linear strand of DNA and proteins in a cell that houses genes . Chro-mosomes are large enough to be seen under a microscope . In humans, all cells other than germ cells usually contain 46 chromosomes: 22 pairs of autosomes and either a pair of X chromosomes (in females) or an X chromosome and a Y chromosome (in males) . In each pair of chromosomes, one chromosome is inherited from the father and one from the mother .
Clarification: A step in the downstream phase of manufacturing a biologic . After the protein product is harvested, which may include remov-ing intracellular proteins from cells, clarification steps separate the protein from cellular debris . Individual proteins are then separated using chromatography methods .
Clinical Trial: A type of research study that evaluates the safety and efficacy of new drugs, medical devices and biologics in human sub-jects . These tests are required by regulatory agencies as a precondition of regulatory clear-ance to market .
Cloning: The replication of a DNA sequence from one organism to create an exact genetic copy; processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning) or organisms .
Codon: A string of exactly three mRNA bases that code for a specific amino acid during translation of mRNA into DNA .
Colony Hybridization: The screening of a library with a labeled probe (radioactive, biolumines-cent, etc .) to identify a specific sequence of DNA, RNA, enzyme, protein or antibody .
Column Chromatography: A type of chroma-tography that uses a column for containing and separating a mixture . It is a commonly used method of purifying proteins .
Combinatorial Chemistry: A discipline in which a large number of new chemicals are created, compiled into a library and screened for potential therapeutic use .
37
Glossary
Cryopreservation: A process whereby cells or whole tissues are preserved by cooling to low subzero temperatures . At these low tempera-tures, any biological activity, including the bio-chemical reactions that would lead to cell death, is effectively stopped .
DNA (Deoxyribonucleic Acid): DNA is a nucleic acid that contains the genetic information used in the development and functioning of all organ-isms . Molecular systems interpret the sequence of these nucleic acids to produce proteins .
DNA Fingerprinting: A technique used to distinguish between individuals of the same species using only samples of their DNA .
DNA Ligase: The enzyme that creates a bond between the ends of single-stranded DNA segments . Where restriction enzymes are the scissors of recombinant DNA technology, DNA ligase is the glue .
DNA Polymerase: An enzyme that attaches complementary nucleotides to a single stranded human DNA .
DNase (Deoxyribonuclease): Any enzyme that catalyzes the breaking up of linkages in the DNA molecule backbone .
Downstream Phase: Involves manufacturing processes including the recovery, purification, formulation and packaging of the protein .
Enzyme-Linked Immunosorbent Assay (ELISA): A biochemical technique to detect the presence of an antibody or an antigen in a sample . It is commonly used to detect infectious agents .
Enzymes: The many proteins produced by organisms to act as biochemical catalysts . Enzymes are the mediators of cell metabolism .
Epidermal Growth Factor Receptor (EGFR): A cell-surface receptor that is activated when bound by epidermal growth factor . Genetic mutations that lead to EGFR overexpression or overactivity have been associated with a number of cancers .
Extracellular Proteins: Proteins found outside of a cell .
Fermentation: A process of growing, or culturing, cells by using enzymes to effect chemical changes .
Fusion Proteins (or Chimeric Proteins): Proteins created through the joining of two or more genes that were originally coded for separate proteins . Translation of this fusion gene results in a single new protein with functional properties derived from each of the original proteins .
Gel Electrophoresis: A technique used for the separation of DNA, RNA or protein molecules by using an electric current applied to a gel matrix . The gel is the medium used to contain, then separate the target molecules . Electropho-resis refers to the use of electricity to move the molecules through the gel matrix . Placing the molecules in wells in the gel and applying an electric current moves the molecules through the matrix at different rates based on their size, charge and/or shape .
Gene: A length of DNA that codes for a particu-lar protein or, in certain cases, a functional or structural RNA molecule .
Generally Regarded as Safe (GRAS): A designation that a substance is considered safe by experts under the conditions of its intended use . Examples are CHO and NS0 cell lines that have GRAS status for therapeutic protein production .
Genetic Engineering: Alteration of the genetic material of cells or organisms in order, for example, to make them capable of making new substances or performing new functions .
Glycosylation: The process by which oligosac-charide units are added to proteins .
Half-life: A measurement of the time it takes for a drug to lose half of its pharmacologic activity or half of its administered amount in the blood-stream or in its target tissues .
Hematopoietic Growth Factors: Protein hormones produced by the body to regulate blood development, affecting the production and maturation of blood-forming cells .
High-Throughput Screening: The process of screening a sample of compounds rapidly and in parallel, then analyzing the results and choosing further screening compounds based on this information .
Hormones: Substances produced by one tissue and conveyed to another through the blood-stream, usually affecting growth or metabolism .
38
Humanized Antibodies: Monoclonal antibodies that have been synthesized by using recombinant DNA technology to avoid the clinical problem of an immune response to foreign substances . Humanized antibodies are produced by merg-ing the DNA that encodes the binding portion of a monoclonal mouse antibody with human antibody-producing DNA . Cell cultures are used to express this recombinant DNA and produce these partial-mouse and mostly human antibodies .
Hybridization: The process of joining two com-plementary strands of DNA or one each of DNA and RNA to form a double-stranded molecule .
Hybridoma: A cell that has been engineered to produce a desired antibody in large amounts . Hybridomas are created by fusing immortal tu-mor cells with antibody-producing B-lymphocyte cells that continuously synthesize identical (or monoclonal) antibodies .
Immortal Cell Line: An established cell line that has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification .
Immunotherapy: Modulation of the immune system to achieve a therapeutic goal . Monoclonal antibodies are a type of immunotherapy .
Interferon: A naturally occurring cell-signaling protein produced by the immune system in response to infections such as viral infections or parasites .
Intracellular Proteins: Proteins found inside a cell .
Investigational New Drug: A drug that has been approved by the FDA for use in human clinical trials .
In vitro: The technique of performing an experi-ment outside of a living organism, in a controlled environment such as in a cell culture or in cells grown in a petri dish .
Ion Channels: Pore-forming proteins that help establish and control the small voltage gradient across the plasma membrane of all living cells . Ion channels are involved in a wide variety of biological processes and are a favorite target in the search for new drugs .
Mass Spectrometry (MS): An analytic technique for determination of the elemental composition of a sample or molecule . It is also used for de-
termining the chemical structures of molecules, such as peptides or proteins . MS consists of ionizing chemical compounds to generate charged molecules or molecule fragments, then measuring their mass-to-charge ratios .
Media: Nutrient-rich substances in which cells are grown .
Messenger RNA (mRNA): A polynucleotide copy of a DNA gene that communicates the code for building a protein to ribosomes so that new proteins can be built .
Microarray: A tool that enables analysis of the levels of expression of genes in an organism or comparison of gene-expression levels .
Monoclonal Antibody: An antibody produced by cells that are all derived from a single antibody-producing cell . Once a cell capable of generating an antibody with desired therapeutic characteristics is selected, laboratory processes are used to clone (make large numbers of) these cells . Since the cells are all identical and are produced by cloning one specific cell in great numbers, they are called monoclonal and can be used to continuously produce identical antibody molecules with these same therapeutic characteristics .
Nanomedicine: The medical application of nanotechnology .
Nanotechnology: The study and creation of systems and devices at the level of molecules and atoms .
Neurotransmitters: Chemicals that are used to relay, amplify and modulate signals between a neuron and another cell .
Non Secreting (NS0) Cells: Mouse myeloma cells that are used frequently in the production of recombinant antibodies .
Nucleotide: The name given to an individual unit of the DNA double helix and RNA . A nucleotide contains one sugar, one phosphate and one base .
Nucleus: The organelle within a living cell that contains genetic material and controls life functions .
Pathogen: A disease-causing agent such as a bacterium or virus .
Pegylation: The process of adding polyethylene glycol to a therapeutic protein, which enables
Glossary 39
Glossary
the therapeutic protein to stay in the body longer .
Peptibodies: Engineered therapeutic fusion proteins with attributes of both peptides and antibodies but that are distinct from each and that can bind to human drug targets .
Peptide Bond: A bond that links together two or more amino acids . A protein is a long chain of amino acids joined together with peptide bonds and therefore is sometimes referred to as a polypeptide .
Peptides: Short chains of amino acids . Poly-peptides, or multiple peptides linked together by peptide bonds, are long chains of amino acids .
Personalized Medicine: Use of the information contained within a patient’s genome, genotype or genomic signature to design and tailor the best treatment plan for that individual patient .
Pharmacodynamics: Studies performed to determine what a drug does to the body .
Pharmacogenomics: The science of under-standing the correlation between patients’ genetic makeup (genotype) and their responses to drug treatment .
Pharmacokinetics: Studies performed to deter-mine what the body does to a drug .
Phosphorylation: The addition of a phosphate (PO4 ) group to a protein or other organic mol-ecule . Protein phosphorylation plays a significant role in a wide range of cellular processes .
Polymerase Chain Reaction (PCR): A method for creating millions of copies of a particular seg-ment of DNA . If a scientist needs to detect the presence of a very small amount of a particular DNA sequence, PCR can be used to amplify the amount of that sequence until there are enough copies available to be detected .
Preclinical Trials (or Studies): Tests that take place in a scientifically controlled setting using cell culture and/or animals as disease models .
Product Pipeline: In the biomedical industry, the term pipeline refers to the number of unique products or processes reported or in develop-ment by a company . Drugs that have entered into clinical trials are said to be “in the pipeline .”
Protein Engineering: A process for isolating and studying proteins and generating proteins with
modified structures by altering the genes that direct their composition .
Proteins: Compounds (chains of amino acids) constituting the ultimate expression product of a gene . Created through the synthesis performed by ribosomes, proteins are the workhorses of living systems, causing chemical processes and changing as their environment changes .
Proteomics: The study of proteins . Proteomics has three major goals: to identify and quantify all the proteins expressed in an organism, to determine the structure and function of each protein and to study the protein-protein interac-tions that affects how one protein interacts with other proteins to control cellular processes .
Receptor (Cell Receptor): A protein molecule, embedded in either the plasma membrane or the cytoplasm of a cell, to which a mobile signaling (or signal) molecule may attach . A molecule that binds to a receptor is called a ligand, and may be a peptide (such as a neurotransmitter), a hormone, a pharmaceutical drug or a toxin, and when such binding occurs, the receptor goes into a conformational change, which usually initiates a cellular response .
Recombinant DNA: A form of DNA that does not exist naturally and is created by combining DNA sequences that would not normally occur together .
Recombinant Proteins: Proteins created by recombinant DNA technology .
Regenerative Medicine: Research into treatments that restore damaged cells with healthy, disease-free cells .
Restriction Enzymes: Enzymes having the ability to cut DNA at a certain location . Scientists use these enzymes to isolate certain types of DNA and place them into new environments . Where DNA ligase is the glue of recombinant DNA tech-nology, restriction enzymes are the scissors .
Reverse Transcriptase: An enzyme used by retroviruses to form a complementary DNA se-quence (cDNA) from an RNA template—usually the genome of the retrovirus . The enzyme then performs a complementary template of the cDNA strand such that a double-stranded DNA molecule is formed . This double-stranded DNA molecule is then inserted into the chromosome
40
of the host cell, which has been infected by the retrovirus .
Ribonucleic Acid (RNA): A molecule similar to DNA, which helps in the process of decoding the genetic information carried by DNA . RNA is a nucleic acid transcribed from DNA; mRNA is then translated into proteins .
Ribosome: The cell structures within which protein synthesis occurs .
RNA Interference: A mechanism that inhibits gene expression at the stage of translation (see translation) or by hindering the transcription (see transcription) of specific genes . This method has been referred to as posttranscrip-tional gene silencing and is an important tool for gene-expression research .
Signal Transduction: The movement of signals from the outside of a cell to the inside . Scientists are attempting to learn more about this process in cancer cells in order to fight the disease .
Single-Nucleotide Polymorphism (SNP): A DNA sequence variation that occurs when a single nucleotide—A, T, G or C—in the genome dif-fers between members of a species . Variations in the DNA sequences of humans can affect how humans develop diseases and respond to pathogens, chemicals, drugs, vaccines and other agents .
Southern Blotting: Transfer by absorption of DNA fragments separated in electrophoretic gels to membrane filters for detection of specific base sequences by radiolabeled complementary probes .
Stem Cell: Undifferentiated, human cells with the ability both to multiply and to differentiate into specific cells .
Thermocycler: A laboratory apparatus, used to amplify segments of DNA via the polymerase chain reaction (PCR) process . The device has a thermal block with holes where tubes holding the PCR mixtures can be inserted . The cycler then raises and lowers the temperature of the block in discrete, preprogrammed steps .
Transcription: The process by which enzymes use the genetic information on a strand of DNA to create a complementary strand of messenger RNA .
Transfer RNA: Molecules that carry amino acids during the process of protein synthesis during translation .
Transformation: The process of transferring DNA from a donor to a recipient cell . Scientists use this process to introduce recombinant DNA to bacteria, yeast and mammalian cell lines .
Transgenic: A term describing an organism containing genetic material from a source other than its parents .
Translation: The process that converts an mRNA sequence into a string of amino acids that form a protein . Translation follows transcription (see transcription).
Upstream Phase: Involves the production of the protein product, most often by using cells (mi-crobial, insect or mammalian) growing in culture .
Vaccine: An agent bearing antigens on its surface that causes activation of the immune system without causing actual disease .
Vector: (1) An organism that serves to transfer a disease-causing organism (pathogen) from one organism to another . (2) A mechanism whereby foreign genes are moved into an organism and inserted into that organism’s genome .
X-ray Crystallography: A method of determin-ing the arrangement of atoms within a crystal, in which a beam of X-rays strikes a crystal and scatters in many different directions . From the angles and intensities of these scattered beams, a crystallographer can produce a three-dimen-sional picture of the density of electrons within the crystal, and the structure of a substance can be determined .
Glossary 41
Timeline of Medical Biotechnology
1950s1952
- Dr . George Gey establishes a continuous cell line taken from a human cervical carcinoma isolated from Henrietta Lacks, who died of the cancer in 1951 . This cell line, containing HeLa cells, is commonly used in medical research .
1953
- Dr . James Watson and Dr . Francis Crick reveal the 3-D structure of DNA .
- Dr . Joseph Murray performs the first kidney transplant between identical twins .
1955
- An enzyme, DNA polymerase, involved in the synthesis of a nucleic acid, is isolated for the first time .
- Dr . Jonas Salk develops the first polio vaccine . The development marks the first use of mam-malian cells (monkey kidney cells) and the first application of cell culture technology to generate a commercial product .
1957
- Scientists prove that sickle-cell anemia occurs due to a change in a single amino acid .
1958
- Dr . Arthur Kornberg of Washington University in St . Louis makes DNA in a test tube for the first time .
- The first automatic protein sequencer, the Moore-Stein amino acid analyzer, is developed .
1960s1960
- French scientists discover messenger RNA (mRNA) .
1961
- Scientists understand genetic code for the first time .
1962
- Dr . Osamu Shimomura of Boston University discovers the green fluorescent protein Aequorea victoria in jellyfish . He later develops
it into a tool for observing previously invisible cellular processes .
1963
- Independent groups in the United States, Germany and China synthesize insulin, a pancreatic hormone .
1964 - The existence of reverse transcriptase is
predicted .- Dr . Samuel Katz and Dr . John F . Enders
develop the first vaccine for measles .
1967
- Dr . Maurice Hilleman develops the first American vaccine for mumps .
1969
- The first vaccine for rubella is developed . It is combined with the measles and mumps vaccines to form the measles/mumps/rubella vaccine in 1972 .
1970s1970 - Restriction enzymes are discovered . These
enzymes cut DNA into pieces and are used for various studies and applications . The restriction enzyme technique becomes a fundamental tool in modern genetic research and opens the way for gene cloning.
- Dr . Har Gobind Khorana synthesizes the first complete gene at the University of Wisconsin–Madison .
1972
- DNA ligase, which links DNA fragments together, is used for the first time .
- The DNA composition of humans is discovered to be 99 percent similar to that of chimpanzees and gorillas .
- The purified enzyme reverse transcriptase is first used to synthesize complementary DNA from purified messenger RNA in a test tube .
1973 - Dr . Stanley Cohen and Dr . Herbert Boyer use
bacterial genes to perform the first successful recombinant DNA experiment, which inserted
Timeline of Medical Biotechnology
42
Timeline of Medical Biotechnology
a recombinant DNA molecule into a cell for replication .
- Dr . Edwin Southern develops a blotting technique for DNA called the Southern blot . It becomes a seminal technology for studying the structure of DNA .
1974 - The U .S . National Institutes of Health (NIH)
forms a Recombinant DNA Advisory Commit-tee to oversee recombinant genetic research .
- The first vaccine for chicken pox is developed in Japan .
1975
- Colony hybridization and Southern blotting are developed for detecting specific DNA sequences .
- The first monoclonal antibodies are produced . Dr . César Milstein, Dr . Georges Kohler and Dr . Niels Jeme develop monoclonal antibody technology by fusing immortal tumor cells with antibody-producing B-lymphocyte cells to pro-duce hybridomas that continuously synthesize identical (or monoclonal) antibodies .
1976
- The NIH releases the first guidelines for recom-binant DNA research .
- Molecular hybridization is used for the prenatal diagnosis of alpha thalassemia .
- Yeast genes are expressed in E. coli bacteria .
1977 - Protocols are developed to rapidly sequence
long sections of DNA .- Genetically engineered bacteria are used
to synthesize the human growth protein somatostatin, marking the first time a synthetic recombinant gene is used to clone a protein . Many consider this to be the advent of the Age of Biotechnology .
- Dr . Robert Austrian of the University of Pennsylvania develops the first vaccine for pneumonia .
1978
- Dr . Herbert Boyer of the University of California, San Francisco, constructs a synthetic version of the human insulin gene and inserts it into the bacterium E. coli, allowing the bacterium to produce human insulin .
- The first test-tube baby, Louise Brown, is born in the United Kingdom .
- The first vaccine for meningococcal meningitis is developed .
1980s1980
- The U .S . Supreme Court rules genetically altered life forms can be patented, opening up enormous possibilities for commercially exploit-ing genetic engineering . The first patent of this nature was awarded to the Exxon oil company to patent an oil-eating microorganism, which would later be used in the 1989 cleanup of the Exxon oil spill at Prince William Sound, Alaska .
- Dr . Stanley Cohen and Dr . Herbert Boyer receive a U .S . patent for gene cloning .
- The first automatic gene machine, or gene- synthesizing machine, is developed in California .
- Founding of Amgen, which will grow to become the world’s largest biotechnology medicines company .
1981
- Dr . Baruch Blumberg and Dr . Irving Millman develop the first vaccine for hepatitis B (not recombinant) four years after the virus is discovered .
- Scientists in Switzerland clone mice . - The first transgenic animals are produced
by transferring genes from other animals into mice .
1982 - The U .S . Food and Drug Administration (FDA)
approves the first biologic—or recombinant protein: Humulin®, Genentech’s human insulin drug produced by genetically engineered bac-teria for the treatment of diabetes .
1983 - Dr . Luc Montagnier of the Pasteur Institute in
Paris isolates the AIDS virus .- Dr . Kary Banks Mullis invents the polymerase
chain reaction (PCR), a technique for multiply-ing DNA sequences . PCR is recognized as the most revolutionary molecular biology technique of the 1980s .
- The FDA approves a monoclonal antibody-based diagnostic test to detect Chlamydia trachomatis.
- The first artificial chromosome is synthesized .- The first genetic markers for specific inherited
diseases are found .
43
Timeline of Medical Biotechnology
1984 - The DNA fingerprinting technique is devel-
oped . When a restrictive enzyme is applied to DNA from different individuals, the resulting sets of fragments sometimes differ markedly from one person to the next . Such variations in DNA are called restriction fragment length polymorphisms and are extremely useful in genetic studies .
- The first genetically engineered vaccine is developed for hepatitis B .
- The entire genome of the HIV virus is cloned and sequenced .
1985 - Genetic fingerprinting enters the courtroom .- Genentech becomes the first biotechnology
company to launch its own biopharmaceutical product .
- Genetically engineered plants resistant to insects, viruses and bacteria are field-tested for the first time .
- Cloning of the gene that encodes human lung surfactant protein is accomplished . This is a major step toward reducing premature birth complications .
- The NIH approves guidelines for performing experiments in gene therapy on humans .
1986 - University of California, Berkeley, chemist
Dr . Peter Schultz describes how to combine antibodies and enzymes (abzymes) to create therapeutics .
- The automated DNA sequencer is invented in California .
- The FDA approves the first monoclonal antibody treatment to fight kidney transplant rejection .
- The FDA approves first biotech-derived interferon drugs to treat cancer . In 1988, the drugs are used to treat Kaposi’s sarcoma, a complication of AIDS .
- The FDA approves the first genetically engineered human vaccine to prevent hepatitis B .
1987 - The FDA approves a genetically engineered
tissue plasminogen activator to treat heart attacks .
- Dr . Maynard Olson and colleagues at Washington University invent yeast artificial
chromosomes, which are expression vectors for large proteins .
- Reverse transcription and the polymerase chain reaction are combined to amplify messenger RNA sequences .
- DNA microarray technology, the use of a collection of distinct DNAs in arrays for expression profiling, is first described . The arrayed DNAs are used to identify genes whose expression is modulated by interferon .
- The FDA approves a diagnostic serum tumor marker test for ovarian cancer .
1988
- Congress funds the Human Genome Project, a massive effort to map and sequence the human genetic code as well as the genomes of other species .
- The first agreement between two companies with parallel patents for cross-licensing of biotech products occurs and becomes the prototype .
1989 - The FDA approves Amgen’s first biologically
derived human therapeutic .- Oil-eating bacteria are used to clean up the
Exxon Valdez oil spill .- A gene responsible for cystic fibrosis is
discovered .
1990s1990
- The first federally approved gene therapy treat-ment is performed successfully on a 4-year-old girl suffering from an immune disorder called adenosine deaminase deficiency .
- The Human Genome Project, the international effort to map all of the genes in the human body, is launched . Estimated cost: $13 billion .
- The FDA licenses the first hepatitis C antibody test, which helps to ensure the purity of blood bank products .
- The FDA approves a bioengineered form of the protein interferon gamma to treat chronic granulomatous disease .
- The FDA approves a modified enzyme for enzyme replacement therapy to treat severe combined immunodeficiency disease . It is the first successful application of enzyme replace-ment therapy for an inherited disease .
44
1992
- The U .S . Army collects blood and tissue sam-ples from all new recruits as part of a genetic dog tag program aimed at better identification of soldiers killed in combat .
- The FDA approve the first genetically engineered blood-clotting factor—a recombinant protein used to treat hemophilia A .
- The FDA approves a recombinant protein to treat renal cell cancer .
- American and British scientists unveil a tech-nique for testing embryos in vitro for genetic abnormalities such as cystic fibrosis and hemophilia .
1993 - The FDA approves a recombinant protein to
treat multiple sclerosis—marking the first new multiple sclerosis treatment in 20 years .
- An international research team, led by Dr . Daniel Cohen of the Center for the Study of Human Polymorphisms in Paris, produces a rough map of all 23 pairs of human chromosomes .
- Two smaller trade associations merge to form the Biotechnology Industry Organization, an international biotechnology advocacy group .
1994
- The FDA approves a recombinant protein to treat growth hormone deficiency .
- Dr . Mary-Claire King at the University of California, Berkeley, discovers the first breast cancer gene, BRCA1 .
- The FDA approves a modified enzyme to treat Gaucher’s disease .
- A multitude of genes, human and otherwise, are identified and their functions described . These include:
•Ob,agenepredisposingtoobesity •BCR,abreastcancersusceptibilitygene •BCL-2,ageneassociatedwithapoptosis
(programmed cell death) •Hedgehoggenes(namedbecauseoftheir
shape) produce proteins that guide cell differentiation in advanced organisms
•Vpr,agenegoverningreproductionoftheHIV virus
- Linkage studies identify genes for a variety of ailments, including bipolar disorder, cerulean cataracts, melanoma, hearing loss, dyslexia, thyroid cancer, sudden infant death syndrome, prostate cancer and dwarfism .
- The FDA approves a genetically engineered
version of human DNase, which breaks down protein accumulation in the lungs of cystic fibrosis patients . It represents the first new therapeutic drug for managing cystic fibrosis in more than 30 years .
1995
- The first baboon-to-human bone marrow transplant is performed on an AIDS patient .
- The first vaccine for hepatitis A is developed . The NIH, the U .S . Army and the Centers for Disease Control and Prevention are significantly involved in the development and clinical testing of the vaccine .
- Scientists at the Institute for Genomic Research complete the first full gene sequence of a living organism (other than a virus) for the bacterium Haemophilus influenzae.
- A European research team identifies a genetic defect that appears to be the most common cause of deafness .
1996
- The Department of Biochemistry at Stanford University and Affymetrix develop the GeneChip, a small glass or silica microchip that contains thousands of individual genes that can be analyzed simultaneously . This marks a techno-logical breakthrough in gene expression and DNA-sequencing technology .
- A group of scientists sequence the complete genome of a complex organism, Saccharomyces cerevisiae, otherwise known as baker’s yeast . The achievement marks the complete sequencing of the largest genome to date—more than 12 million base pairs of DNA .
- A new, inexpensive diagnostic biosensor test is developed to allow instantaneous detection of a toxic strain of E. coli, the bacteria responsible for many food-poisoning outbreaks .
1997 - The first human artificial chromosome is cre-
ated . A combination of natural and synthetic DNA is used to create a genetic cassette that can potentially be customized and used in gene therapy .
- The FDA approves a recombinant follicle- stimulating hormone to treat infertility .
- The FDA approves the first bloodless HIV-anti-body test that uses cells from patients’ gums .
- Scientists at the Institute for Genomic Research sequence the complete genome of the Lyme
Timeline of Medical Biotechnology 45
Timeline of Medical Biotechnology
disease pathogen, Borrelia burgdorferi, along with the genome for the organism linked to stomach ulcers, Helicobacter pylori.
- Scientists at the University of Wisconsin– Madison sequence the E. coli genome .
- The FDA approves the first therapeutic antibody to treat cancer in the United States . It is used for patients with non-Hodgkin’s lymphoma .
1998 - Human skin is produced for the first time in
the lab .- Two research teams culture embryonic stem
cells . Embryonic stem cells are used to regen-erate tissue and create disorders mimicking diseases .
- Scientists at the Sanger Institute of the Washington University School of Medicine in St . Louis sequence the first complete animal genome for the Caenorhabditis elegans worm .
- A rough draft of the human genome map is produced, showing the locations of more than 30,000 genes .
- The first vaccine for Lyme disease is developed .- The FDA approves a novel monoclonal anti-
body to treat Crohn’s disease .- A monoclonal antibody therapy used against
breast cancer has favorable results, heralding a new era of treatment based on molecular targeting of tumor cells .
- Approval of the Her-2 inhibitor for the treatment of breast cancer patients who have tested positive for the Her-2 mutation brings personalized medicine to oncology .
1999 - The complete genetic code of the human
chromosome is deciphered .
2000s2000 - Scientists at Celera Genomics and the Human
Genome Project complete a rough draft of the human genome .
2001
- Science and Nature magazines publish the hu-man genome sequence, making it possible for scientists all over the world to begin research-ing new treatments for diseases that have genetic origins, such as cancer, heart disease, Parkinson’s and Alzheimer’s .
2002 - An era of very rapid shotgun sequencing of
major genomes is completed . Included are the mouse, chimpanzee, dog and hundreds of other species .
2003 - Celera and NIH complete sequencing of the
human genome .
2004 - The FDA approves the first monoclonal anti-
body that is an antiangiogenic, inhibiting the growth of blood vessels—or angiogenesis—for cancer therapy .
- The FDA clears a DNA microarray test system, which aids in selecting medications for a variety of conditions . This is an important step toward personalized medicine .
2006 - The FDA approves a recombinant vaccine
against human papillomavirus, which causes genital warts and can cause cervical cancer .
- Scientists determine the 3-D structure of the human immunodeficiency virus, which causes AIDS .
2007 - Scientists discover how to use human skin
cells to create embryonic stem cells .
2008 - Chemists in Japan create the first DNA mol-
ecule made almost entirely of artificial parts . The discovery can be used in the fields of gene therapy .
- Dr . Craig Venter and his team replicate a bacterium’s genetic structure entirely from laboratory chemicals, moving a step closer toward creating the world’s first living artificial organism .
46
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