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2004 11:41 Uhr Seite 9
Jens-Peter Mendelsohn
Planning and Construction of Complete Pharmaceutical Plants
Biotechnology Plant for
Insulin Production
About 800,000 people in Germany alone must use insulin every
day to control their blood sugar level. Because of the speedy
progress of genetic technology in the past 30 years, it has become
possible to make almost unlimited amounts of human insulin and
insulin analogs with consistently high quality using genetically
modified microorganisms. The most modern biotechnological
production plants are needed to meet the demand for innovative
analogs, such as glargin insulin.
Planning and construction of pharmaceutical plants is
a relatively young field of business for Linde Plant
Construction, located in Dresden at Linde-KCA (LKCA).
Even so, LKCA has, over the past ten years, established
itself as a leading bidder in the design, planning and
construction of pharmaceutical and biotechnological
plants. The latest reports are among others which confirm
that: the Bayer AG pharmaceutical pilot plant in
Elberfeld, which started up in October 2000, and the
Figure 1: Lantus
Aventis at the Höchst
Industrial Park.
Linde Technology I 1/ 2004
new F. Hoffmann-La Roche AG “Kilolabor” in Basel, are
both plants for synthesis of innovative pharmaceuticals.
LKCA obtained its first contract, as general planner
for construction of a new biotechnology plant, in April,
2000. The plant, for Aventis Pharma, Frankfurt, is to
produce glargin insulin, a genetically engineered insulin
derivative with depot action. LKCA turned the plant
over to Aventis only 29 months later, after successful
completion of the start-up activities, for continuation of
special performance tests by Aventis, and for the
beginning of Validation (Figures 1 and 2).
A stand-alone plant
on the ‘green-field’
The plant is an independent complex consisting of
four buildings among which the production and auxiliary
operations are distributed on the basis of their require-
ments for classified cleanrooms and for areas and
processes with explosion protection. All the subplants
are supplied through a central connecting road. The road
serves the flow of both personnel and materials. It
also connects the central main building to the offices,
conference rooms, laboratories and the real heart of the
plant, the central control room. There are also
storage tanks as well as the entire infrastructure with
lines from the plant to medium supply and disposal at
the Höchst Industrial Park (Figures 2 and 5). The plant
produces about 1,700 kg of glargin insulin every year.
The total investment is around 200 million euro. Because
its amino acid sequence has been altered by genetic
engineering, this insulin derivative has depot action
for 24 hours in the body. Glargin insulin was developed
independently by Aventis, which was the client for
this project. The product is made in the subplants for
fermentation, processing, purification and end-product
treatment. The production complex also includes supply
and disposal installations (propanol distillation plant,
auxiliary material preparation, wastewater disposal, tank
storage and a warehouse for packaged goods).
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After disintegration, the specific heavier fusion protein is

Production in five steps
The plant for producing biosynthetic glargin insulin is
organized into these five processing sections:
■ Fermentation
■ Processing 1
■ Processing 2
■ Purification, and
■ End product treatment.
The steps in the process for producing glargin insulin
differ from those for producing rapid-acting insulins
essentially in purification and end product treatment.
The fermentation and processing stages use similar
production technology.
In the fermentation stage genetically modified
Escherichia coli bacteria are cultured and grown in
successive fermenters with increasing capacities. They
are stimulated to produce a fusion protein by adding
an inducer. When the fusion protein has been formed,
the bacteria are killed with a disinfectant. This part of
the process – culture and killing of genetically modified
bacteria – is covered by the Genetic Technology Act,
and is the genetic engineering part of the process.
After killing, the bacterial suspension is concentrated
by centrifugation during processing, and then disintegrated.
separated from the other cell components by continuous
centrifugation, washed twice with water, and isolated.
The “folding” of the molecule, in which the fusion protein
folds into a “native spatial structure” takes place in
the next step of the process. That is accomplished by
dissolving the fusion protein in an aqueous urea
solution in the presence of cysteine. Byproducts are then
precipitated by a shift in the pH and separated by
centrifugation.
In the subsequent purification process, the
glargin insulin is freed of urea and inorganic salts by
adsorption-desorption steps and simultaneously
concentrated. The desorption is done with an aqueous
solution of propanol. The crude intermediate product is
then purified chromatographically in two process steps.
Then it is crystallized at high purity, filtered off by
suction, and stored temporarily as moist crystals.
The high-purity glargin insulin crystals are dissolved
again, crystallized and freeze-dried to get larger
homogeneous batches. The end product treatment is
done in Class C cleanroom conditions because there
are limits for the bacterial counts and endotoxins
(“pyrogens”) in the dried material. The plant, which only
produces the solid pharmaceutical, not the finished
medicinal product, had to be approved not only under
the Genetic Technology Act but also under the Federal
Environmental Protection Act.

Figure 2: View of the

tank storage, with
the propanol distilla-
tion plant and the
fermentation build-
ings (right).

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Diabetes mellitus

Diabetes mellitus is one of the most
common and significant metabolic
diseases in humans. It can be caused
by malfunction of the pancreas,
which produces too little insulin, or
by insulin resistance of the body cells.
The result is that the concentration
of sugar in the blood is no longer
normal and must be corrected by
medications. Although the pancreas
of a healthy person produces 2 mg
of insulin a day, a diabetic must get
about 1.5 mg of insulin per day
externally. In Germany alone, about
800,000 people must take insulin
daily to adjust their blood sugar
levels.
According to an estimate by the
World Health Organization (WHO)
there are at least 120 million
diabetics in the world today, includ-
ing undiagnosed cases. More than
8 million patients are being treated
with insulin in the industrialized
countries alone. The worldwide
insulin consumption is around five
to six tons per year. It is predictable,
too, that this requirement will
increase substantially in the future,
in view of the worldwide increase
in population and changing social
structures.

Exacting production process
All the steps of the process use deionized and purified
water. Its microbiological and physical-chemical quality
are monitored routinely (“Purified water”). Ozone is
removed from the water with UV light before it reaches
the points where it is used. The consumers are supplied
with process water through a ring piping system.
“Pyrogen-free purified water” produced by ultrafiltration
is used In the last steps of the process (purification and
end product treatment).
In addition, all the raw materials and auxiliary
materials (solvents, acids, bases, salts, buffers, etc.) are
of pharmaceutical quality or meet internal plant
specifications, and are checked regularly. The solvent,
propanol, is distilled after use and returned to the
process. The nitrogen used for inerting is purified through
a HEPA (high-efficiency particulate air) filter before use
to prevent contamination by particles.
As a rule, all the stages of the process are carried
out in closed tanks with solid pipe connections. Manual
handling of the product is reduced to a minimum. The
material used for the tanks and connecting pipes are
corrosion-resistant stainless steel with defined surface
roughness (internally electropolished for critical process
steps). This assures good and complete cleaning. These
cleaning processes have a critical part in the planning of
pharmaceutical plants to assure the required sterility and
constant product quality. They are generally established
by the client in the corresponding specifications.
Process monitoring
and documentation
The heart of the glargin insulin production plant is a
high-capacity process control system, which processes
about 18,000 items of input and output information
and monitors and controls the entire process.
All quality-related instruments were calibrated when
they were put into service. This calibration is repeated
regularly at appropriately established intervals. In-process
samples of the intermediate products are also tested in
the laboratory to make sure that their quality meets
specifications (Figure 4).
All the process and analytical data, either from
routine operations or from deviations, are stored in the
process control system or in the laboratory information
management system (LIMS). Then the specific process
data are supplemented with the general operating
data and printed out either in partial lot records or a full
lot record, along with the operational data.
Conceptual design –
the key to success
This project began with a process which Aventis
developed and tested at pilot plant level, which had
to be converted to a full-scale plant. Utilization of the
structure of a similar project for insulin production,
which had already been accomplished, was a significant
boundary condition. In this phase, the plant designers
were expected to apply their experience and know-how
to work out the technological problem and to participate
actively in the design process, especially with respect
to process optimization, scaling up and the defined
production concept.

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Figure 3: The 3D
design model and the
structural plan of the
plant for producing a
genetically engineered
insulin derivative
with depot action.

Tank Storage
Processing
Fermen-
tation
Laboratory and Administration

Connecting Structure – Central Supply Route

Main Building

High Purification Warehouse and Supply

Purification and End Product
Treatment

The major emphasis in planning is that of making sure ■ functional layout

that the product is produced under reproducible ■ building concept
conditions (with respect to product quality and ■ master plans for beginning-to-end quality
validation). LKCA set up a team of 12 employees assurance, from design to start-up
specifically for the project. They worked together with ■ plant design, connected to the existing
the Aventis project team for three months following infrastructure.
April 2000, under a preliminary contract, to develop the
initial plant concept. This process was essential so that the documents
needed for the construction contract and for the Federal
In this planning phase, it was important to bring Environmental Protection Act contract could be produced
the process technology into harmony with these as quickly as possible on an assured design basis and
requirements: submitted to the authorities responsible.
■ building architecture
■ cleanroom classification and HVAC The conceptual design produced in that manner
(heating, ventilation, air conditioning) was based on:
■ logistics and storage ■ Layout studies of critical plant areas
■ information technology and process control (e. g., recrystallization) with respect to
■ functional descriptions as the basis for structure, GMP requirements, material
the formulating operation handling, cleaning, and serviceability
■ product quality and GMP (Good Manufacturing ■ layout designs for medium supply and
Practice) concepts. disposal, electrical engineering, measurement,
and control technology, as well as HVAC
Plant-oriented design planning was established, ■ logistical concepts with respect to material,
starting from the basic data determined and specifi- personnel and product flow.
cations of process-oriented design principles (Figure 3):

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Figure 4: Quality
monitoring by
offline-testing in the
analytical laboratory
in the main building.

Insulin glargin (Lantus®)

Glargin insulin is a peptide hormone
analog of insulin, produced by
genetic engineering. Like human
insulin (NPH insulin), it controls the
blood sugar balance in humans,
but has a long-term action profile.
Because of that, a diabetic needs
only one insulin injection per day.
That is a definite improvement from
the patient’s viewpoint. This makes
glargin insulin a product that is
tailor-made for diabetics. Together
with the classical human insulin
(with a relatively fast action profile),
it rounds out the patient’s require-
ment. The active pharmaceutical
agent, glargin insulin (solid, non-
sterile) is dissolved in the finished
medicinal production and supple-
mental materials are added. The
product is filled into cylindrical
ampules, and marketed under the
tradename Lantus®. Glargin insulin,
like human insulin, is a protein
molecule, which reduces the blood
sugar content to the normal value of
about 70 to 120 mg/100 ml.
In current treatment of diabetes,
human insulins play an important
role in the basal supply of insulin for
the body. NPH insulins exhibit a
distinct maximum action after about
4 to 7 hours, and have an effective
duration of about 12 to 16 hours. In
contrast, glargin insulin does not
have any distinct maximum activity,
instead having a constant flat action
profile over 24 hours.

Serum insulin Previous NPH insulin

level

Now: LANTUS®

Time
The difference in the action profile between glargin insulin and NPH insulin.

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Figure 5: View to the

southeast of the com-
plete Aventis Lantus
plant at the Höchst
Industrial Park, with the
central office building in
the foreground.

The total investment is about 200 million euro. The

centerpiece of the glargin insulin production plant is a
high-capacity process control system, with which about
Successful plant start-up 18,000 pieces of input and output information are
processed, and which monitors and controls the entire
Aventis received approval for construction promptly in course of the process.
December 2000, so that the groundwork could be
started in January 2001. Laying of the cornerstone was
celebrated jointly on 4 April 2001. Then it was possible
to complete the rough construction work, an important
project milestone, in August 2001. The mechanical
preparation was done building-by-building, oriented to The Author
the course of the process, until August 2002. After
finishing the assembly, we could then begin the start-up
in the same sequence.
The plant is distinguished by a high degree of Jens-Peter Mendelsohn
complexity. It is a large-scale computer-controlled plant.
Just one single person at the control room computer Jens-Peter Mendelsohn, a gradu-
screen could manage it. It does require about 6,500 ate engineer, has been manager
control points (connections to the process control of the Development Department
system). The start-up of the entire process control system in the area of Pharmaceutical
ran smoothly. By February 2002 the technology was Plants since 2003. He started at
available to start operations of the individual sections Linde in 1980, as manager of plant design, and,
of the plant. after 1987, he coordinated use of CAD technology
for pharmaceutical plants. Since 1990 he has been
responsible for the Central Engineering Technology
Department. From 1995 to 2002 he directed various
major projects, including the construction of the
new Aventis insulin plant.
Abstract
Within ten years, Linde-KCA-Dresden GmbH has
established itself as a leading bidder in the area of
pharmaceutical plants. In just 29 months, under a
contract with Aventis, Linde built a plant to produce
genetically engineered insulin (glargin insulin). The
plant produces bout 1,700 kg of glargin insulin annually.

14 Linde Technology I 1/ 2004