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KB-0002-Jun08
ISPE
Fundamental
Knowledge Brief
Biotechnology Basics
The information
contained in this Knowledge
Brief was adapted from
training materials as part of
the ISPE Training Course on
Biotechnology Basics,
taught by Jeff Odum.
Introduction
Increasingly, the pharmaceutical industry
has undergone a shift in emphasis from
traditional chemical based medicines to
those involving large molecules. The
large molecule medicinal is a direct
result of the major role of biotechnology
in our industry.
“Biotechnology is a combination
of advances in our
understanding of molecular and
cell biology and human
genetics, and how the human
immune system fights disease.”
This modern definition is changing
rapidly as the technologies continue to
develop and advance within the industry.
For obvious and good reason, a major
application of biotechnology is toward
human health care, including: the
detection and treatment of diseases;
human growth; and vaccines.
Biotechnology holds tremendous
potential in other applications that have
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major economic and social impact in our
society, such as agriculture, food
processing, fuels, and wastewater
management. The focus of this
Knowledge Brief will be on
biotechnology’s application toward
human therapeutics. The general
information provided here is drawn from
ISPE training courses that are focused
on biotechnology and biomanufacturing
processes.
The purpose of this Brief is to provide
basic concepts explaining the science of
biotechnology and how science and
process are combined to lead to the
manufacture of a human therapeutic
product. More detailed information can
be found in ISPE publications and in
select ISPE training programs available
on-site or on-line.
The Science of Biotechnology
Cells are the fundamental working units
of all living things. Within the cell are all
the instructions needed to direct the
cells activities. These instructions are
stored in deoxyribonucleic acid (DNA) in
the nucleus of cells.
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Biotechnology Basics
only tiny quantities can be extracted
from human tissue. A significant and
worthwhile goal of biotechnology is the
production of sufficient amounts of high
quality human proteins. The structure of
the DNA molecule is identical among all
living things, from an amoeba to a killer
whale, from a blade of grass to a
redwood tree. This is what makes
recombinant DNA technology – the
transplanting and combining of genetic
information, one organism to another –
possible. The term recombinant DNA
technology is more popularly known as
genetic engineering.
Bioprocess Basics
Now let’s take a look at how the science
and the process integrate to lead to the
manufacture of a human therapeutic
product.
Figure 1. Human Genome Program, U.S. Department of Energy, Genomics and It’s Impact on
Medicine and Society: A 2001 Primer, 2001.
DNA is a long molecule that is
composed of a sugar, phosphate, and
one of the following four nitrogen bases:
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Adenine (A)
Thymine (T)
Guanine (G)
Cytosine (C)
A sugar, phosphate, and base together
constitute a nucleotide. These
nucleotides pair up into strands that twist
together to form a double helix. A
segment of DNA, in conjunction with
A process is defined as any operation or
series of operations by which a particular
objective is accomplished. In this case,
the objective is the production of a
biotech drug product.
some proteins, is a chromosome.
Each chromosome contains many
genes. Genes contain instructions for
making proteins. Proteins are the major
structural and regulatory molecules
essential for life. Proteins act alone or in
complexes to perform a particular
function in the body.
Human proteins are valuable in treating
disease. For example, diseases caused
by a protein deficiency can be treated
with the human protein itself. However,
The DNA Code
In Figure 2, the “basic” biotech process
steps are shown as upstream and
downstream operations. Upstream
operations can be thought of as cell
growth and separation. Downstream
operations are product purification and
filling. Almost every biotech process can
be sub-divided into these two basic
elements.
The basic unit operations are shown in
Figure 3. These unit operations
represent the “standard” operations that
most companies implement in their
manufacturing operations.
The basic biotech process elements are:
The DNA alphabet is A, G, C, and T. The sequence of these four letters
determines DNA information. A DNA sequence is actually a string of three-letter
“words.”
AGCTTCCGATCGGTA actually reads: AGC TTC CGA TCG GTA.
Each three letter sequence, or condon, specifies one of 20 amino acids. Amino
acids are the subunits of protein. AGC is the code for the amino acid Serine and
TTC is the code for Phenylalanine. No matter where they are found, the condons
are always the same for the same amino acid.
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Bioanalysis
Fermentation
Separation
Purification
Filling
Bioanalysis
Nearly every process conducted in a
biopharmaceutical company requires
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ISPE Knowledge Brief
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Biotechnology Basics
number of different options that
companies have as a method of
breaking down cellular material:
• Nonmechanical
- Freezing
- Detergents
- Enzymes
• High pressure
- Centrifugation
• Homogenization
• Mechanical grinding
Most common in use today is
centrifugation. Centrifuges are one of
the most common equipment items used
for cell disruption. A GMP model can
cost well over $500,000 (US).
The downstream operations begin by
separating the “good” from the “waste” in
the product materials. Separating is
nothing more than a filtration operation
to accomplish this activity.
Figure 2.
“back up.” The analysis phase of
manufacturing is critical as proof of the
drug’s safety, purity, and efficacy.
Analytical Methods
Back up regulatory submissions.
• Support pre-clinical and clinical
studies.
• Monitor environmental conditions
during manufacturing.
• Monitor quality of the manufacturing
process.
Cell culture is a specific type of
fermentation that involves the process of
taking cells from living organisms and
growing them under controlled
conditions. Cell culture is part of the
upstream processing operations; literally
engineering and growing the cell line to
be used to manufacture the drug
product.
The separation steps are:
Once fermentation is complete, the
desired product must be recovered,
separated out, and purified.
Purification
Fermentation
Separation
Fermentation is the process by which
living cells obtain energy through the
breakdown of glucose and other
molecules. Fermentation refers to the
large-scale cultivation of
microorganisms.
Recovery is the separation of crude
product from microbial mass and other
solids and liquid medium, to prepare it
for purification.
Figure 3. Basic Processing Steps.
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Product recovery usually requires some
type of cell disruption. There are a
• Extraction and precipitation
• Filtration
- Microfiltration
- Ultrafiltration
Filtration skids are highly automated and
can cost in excess of $250,000 (US).
The purification steps are:
• Chromatography
- Gel filtration
- Ion exchange
- Hydrophobic interaction (HIC)
- Affinity
The purification steps are high risk and
very costly to perform. Formulation of
the drug is a critical step that is very
important to the patient.
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Biotechnology Basics
The required protein must be modified to
a stable, sterile form that can be taken
by the patient. Remember, proteins
cannot be taken as pills because they
are broken down in the stomach. Until
recently, biotech products were
exclusively sterile injectibles. Now,
inhalation and transdermal delivery
options give patients greater flexibility.
Filling
Filling is the process of putting the drug
product into a container. Two general
categories of filling are:
• Bulk
• Final
Bulk filling is defined as the placement of
larger quantities (5L-100L) of product
into containers for shipment/storage.
Some examples of containers are:
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Vials
Ampoules
Syringes
Dental cartridges
Final filling is defined as the placement
of drug product into its final container/
closure system.
The majority of production facilities
produce product in bulk. Many
companies ship their bulk to contract
filling firms.
Conclusion
This document is a basic review of the
general principles of biotechnology and
biotech processing. As such it is
intended to serve as a primer on this
highly sophisticated area of science and
technology. For more detailed
information, you can search ISPE
articles and publications on this subject,
including the ISPE Baseline®
Pharmaceutical Engineering Guide
Series: Volume 6: Biopharmaceutical
Manufacturing Facilities. For the most
current information and up-to-the-minute
discussions on biotechnology and
biotech processing, visit our
Biotechnology Community of Practice on
the ISPE Web site.
About the Author
Jeffery Odum is president of
NCBioSource USA and ISPE’s North
American Continuing Education Advisor.
He has been involved in the
biopharmaceutical industry for more than
20 years with expertise in engineering
design, construction, and plant
operations. Prior to founding
NCBioSource, he was a successful
market leader for an international
engineering design firm specializing in
biopharmaceutical manufacturing facility
design and construction. His experience
includes design and construction of many
of the industry’s major manufacturing
projects, as well as consulting roles for a
number of the global biotechnology
industry leaders. Odum is a nationally
recognized author and speaker with
industry insight in the areas of regulatory
compliance, facilities and process design,
and project management for
biopharmaceutical companies. He has
been a Member of ISPE for more than 15
years and has been a presenter and
course leader for more than 25
professional training and education
sessions within the industry. Odum is a
past chairman of ISPE’s North American
Continuing Education Committee (NAEC)
and the ISPE Training Committee. As
North American Continuing Education
Advisor, he works closely with the NAEC
and the Director of Continuing Education
on development of the society’s North
American conferences. He is also a
Member of ISPE’s Technical Training
Staff. In 2002, Odum received the
Society’s prestigious Richard B. Purdy
Award for Outstanding Achievement
within the biopharmaceutical industry.
Odum is the author of more than 25
published works on many critical issues,
including process improvement and
execution to meet regulatory guidelines
issued by the FDA and other international
regulatory bodies. He was also one of the
lead chapter authors for ISPE’s
Biopharmaceutical Manufacturing
Facilities Baseline® Guide. Odum
graduated from Tennessee Technological
University with a degree in mechanical
engineering and received his Masters
Degree in engineering from the University
of Tennessee-Knoxville.
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© Copyright ISPE 2008. All rights reserved.