Advances in Sensor Technology Improve

Biopharmaceutical Development
by Amber Ratcliff and Carol PreisigMonday, April 1, 2013 9:00 am

Todays
biomanufacturing
operations
require
constant
management
of
biopharmaceutical process attributes throughout process development and production.
Continuous online measurements of pH, dissolved oxygen (DO), oxidationreduction
potential (ORP), and conductivity (Figure 1) allow real-time industrial process monitoring
and adjustment. These functions are crucial to process improvement studies and
accurate, reliable manufacturing of high-quality products.

Figure 1: ()

In the pharmaceutical industry, it is

extremely valuable to see how an attribute
changes with time and correlate that
change with parts of the process, says L.
Harry Lam, PhD, a biopharmaceutical
manufacturing industry expert. It is imperative to have effective equipment that
provides reliable measurements.

LEVEL: BASIC
Accuracy, repeatability, response time, and detection limits are important, and so are
easy installation, calibration, and maintenance. Until the past few years, sensor
technology was not keeping pace with improvements in cell engineering processes,
which have been scaled up and modernized considerably. But three recent advances
optical DO sensor technology, smart sensors, and wireless technology are
improving data quality and the ease with which information is obtained and managed.
Here we describe those advances and present two examples of wireless smart sensors
in use: at an innovative, large-scale biopharmaceutical manufacturing plant and a
scaled-down simulator for studying the impact of industrial-scale inhomogeneities on
microbial cultures in a laboratory setting.

Dissolved Oxygen Sensor Technology Advances

New optical DO process sensor technology was introduced for the biopharmaceutical
industry in 2007 (1). In the classical amperometric (electrochemical) procedure as
described by Clark, oxygen diffuses through a membrane and induces a chemical
reaction with the electrolyte behind it (1). That creates a voltage differential proportional
to the amount of oxygen present in the system.
Optical measurements are based on fluoresecnt quenching of a luminophore when
oxygen is present. The intensity and phase shift of emitted light depends on the amount
of oxygen present. That light is trapped and measured. The light system is located
behind the membrane cap and protected by sapphire glass.
The new sensors replace traditional Clark cells, which were based on electrochemical
measurement within each sensor itself. Because of pressure effects on the fluid-filled
membrane cap, however, classical amperometric sensors could give inaccurate
readings when they were exposed to pressure and temperature changes. That makes it
difficult to assess the status of a culture, triggers alarm systems unnecessarily, and can
disrupt daily operations. By contrast, optical DO sensors have more rugged sensor caps
that easily handle broad performance ranges, accommodating temperatures from 10
C to 80 C, pressures up to 12 bar (174 psi), and pressure spikes up to 80 bar (Table
1).
Table 1:Technical specifications for the Hamilton VisiFerm DO optical DO sensor.

Table 1:Technical specifications for the

Hamilton VisiFerm DO optical DO sensor.
()

Figure 2 illustrates the performance difference between amperometric and optical DO

sensors. Figure 2, TOP, presents the readings of one amperometric and three optical
sensors for a stop-flow process in which the sudden stoppage of flow sends a shock
wave (pressure hammer) through the liquid. That pressure hammer transfers through
the electrolyte-filled membrane of the amperometric sensor, causing a massive
disturbance of the oxygen equilibrium on both sides of that membrane. Optical oxygen
sensors are resistant to pressure hammers because they lack such a fragile membrane
and electrolytes inside the membrane cap. As seen in Figure 2, TOP, the amperometric

sensor triggered an oxygen-level alarm several times during the study, whereas the
three flow-dependent optical sensors provided consistent readings.

Figure 2: ()

Optical DO sensors are also better suited

to modern biopharmaceutical processes
that require steaming-in-place (SIP),
cleaning-in-place (CIP), and autoclaving.
By comparison, amperometric sensors
recover slowly after cleaning because they
rely on requilibration of electrolytes across
a fragile membrane, and their signal
quality
deteriorates
with
repeated
sterilization. Slow recovery extends downtime and can lead to waste if sensors are not
fully responsive before restarting a process. With no electrolytes that would require
polarization time, optical sensors retain their stability over a number of sterilization
cycles (Figure 2, BOTTOM).
Smart Sensors
Bioprocess operation performance has also been hindered traditionally by difficulties in
transmitting information to and from sensors. Recent introduction of so-called smart
sensors facilitates both sensor and data management.
A smart sensor contains a memory chip embedded in its electronic circuitry to store
identification and calibration information. The memory chips provide several benefits.
They eliminate the need for in-line calibration, reducing system downtime. They reduce
incorrect estimations of sensor life, lowering the instance of batch failures attributable to
sensor failure. And they eliminate the need for extensive manual documentation of
sensor
performanc
e to meet quality system reporting requirements, which in turn reduces overall
administration costs.
Smart sensors self-monitor their own measurement performance. An intrinsic quality
indicator will predict a failing sensor before a bioreactor run starts, reducing the risk of
in-process failure and maximizing the useful life of each sensor instead of relying on a

set replacement schedule. Out-of-range results are reported immediately, triggering

alarms and prompting operator action as appropriate. Figure 3 is an example of a
sensor quality readout.

Figure 3: ()

Because smart sensors store calibration

data, precalibration and configuration are possible right in the laboratory. Costs
associated with installation and downtime are reduced by more than half with smart
sensors (Figure 4). Smart sensors improve time to market, allowing less complex setup
and faster qualification, says Lam.

Figure 4: ()

Finally, smart sensors report identification

and calibration information electronically.
This provides for better quality system
adherence and saves time that was
previously
spent
on
manual
documentation.
Wireless Technology
Nearly everyone today is familiar with
wireless information transfer in some form.
The same technology that enables the
function of cellular phones and other
common devices can also be used to create wireless sensor networks in pilot
laboratories and in new bioproduction facilities. Such technology reached the
biopharmaceutical process sensor industry with Hamilton Companys introduction of the
Arc sensor system.

Wireless adapters for sensors are available for a broad range of pH, DO, ORP, and
conductivity measurement applications. Wireless sensors feature a built-in
microprocessor that communicates with analog (420 mA) and digital Modbus
interfaces. (Modbus is a standard serial communications protocol commonly used in
connecting industrial electronic devices. It was developed with industrial applications in
mind.) Direct connectivity eliminates the need to send information through costly
transmitters, and the wireless signals are more robust and reliable than the low currents
produced in traditional measurement systems. Arc sensors contain smart sensor
technology and can be precalibrated and configured by the vendor. In addition, on- and
offline sensor management delivers extended sensor life and increases process
accuracy in challenging bioproduction environments.
In the Hamilton system, an Arc Wi sensor adapter and an Arc View handheld unit
enable wireless management and calibration of multiple sensors using one device. The
handheld unit (Figure 5) provides a local display configured with Arc Wi wireless
communication, which enables users to move around a facility and monitor data from
multiple sensors, communicating with up to 31 of them simultaneously. Figure 6 shows
a manufacturing plant installation with an Arc sensor unit in active transmission mode.

Figure 5: ()

Figure 6: ()

Improving Data Quality with Wireless Sensor Technology: In addition to the more
obvious benefits of monitoring data from multiple sensors using a handheld device,
wireless sensor technology also provides a marked improvement in data quality.
Wireless capability eliminates not only the fixed transmitter and connective cabling
between it and the sensors, but also other hardware that can cause signal interference.

Poor sensortransmitter connections, breaks and shorts in existing cabling, and

sensitivity to electromagnetic fields from nearby motors are just a few well-known
interference sources that can frustrate plant operators. Figure 7 illustrates the dramatic
improvement in data quality that is enabled by removing cables as a source of
interference.

Figure 7: ()

Beyond the advantages of convenience,

flexibility, cleanliness, and elimination of
wiring costs, says Lindsay Leveen, a
bioprocess industry strategist, removing
intrusive wiring ensures quality by
reducing penetrations into the system.
Lower Costs of Smart, Wireless Sensors:Table 2 lists the costs for installing and
maintaining wireless smart sensors using the Hamilton intelligent Arc VisiFerm DO (an
optical DO sensor) as an example. For operation of a single-sensor system over a twoyear period (including installation and one additional year of maintenance), a smart
wireless sensor costs about the same as its traditional hard-wired counterpart. Wi
reless technology provides significant savings when multiple sensors are needed largely
by replacing individual transmitters with a single, less expensive, wireless set-up. For a
five-sensor installation, using a wireless installation with one year of maintenance costs
29% less than would multiple separate traditional transmitters.
Table 2:Cost comparison (in US dollars) for single- and five-sensor installations
of optical DO sensors with and without wireless capability; the second-year cost
is for a replacement cap.

Table 2:Cost comparison (in US dollars) for

single- and five-sensor installations of optical DO sensors with and without wireless
capability; the second-year cost is for a replacement cap. ()

Case Studies
A Modern, 750-L Fermentation Plant Including a Cross-Flow Unit: When planning a
modern, 750-L fermentation plant including a cross-flow unit, a biopharmaceutical
manufacturer needed the best available measurement technology to meet its regulatory
requirements. After testing Hamiltons new smart Arc system, the plant engineer found
that it met the companys needs for efficiency, compactness, reliability, and precise
process controls (Figure 8).

Figure 8: ()

Hamilton Arc sensors were installed in the

plant, which contains nine measurement
points for pH, DO, and conductivity. The
intelligent wireless sensors provide measurement value, sensor quality information,
operating time, CIP/SIP count, product numbers, and calibration data (Figure 9).

Figure 9: ()

Wireless Smart Sensors in a University

Setting for Research in Simulated,
Industrial, Large-Scale Processes: In a bioprocess engineering laboratory, the
combination of a plug-flow reactor (PFR) and common stirred-tank bioreactor (STR) are
used to study the impact of inhomogeneities on Bacillus subtilis cultivations. To
accurately mimic large-scale conditions, operators needed reliable measurement of pH
and DO along their PFR (Figure 10). They chose to use Hamilton smart Arc sensors,
such as the VisiFerm DO Arc 120 and the Polilyte Plus Arc 120 pH models. In Figure
10A, the numbers correspond as follows: (1) recirculation; (2) 11-mm sampling ports; (3)
PFR; (4) insulation; (5) STR; (6) bioreactor outlet; (7) PFR feed; (8) pump; and (9)
optional PFR aeration.

Figure 10a: ()

Figure
10b: ()

A PFR also known as a continuous tubular reactor

(CTR) is a continuously flowing reaction system. It
can be perceived as a tube in which the reaction
medium flows in distinct packets (plugs) of varying
composition in an axial direction. PFRs are typically
used for continuous production. Such a model
system was used to examine the metabolic response
of nonsporulating B. subtilis mutant strains to
oscillating substrate and oxygen availability. When air
was supplied from the bottom (Figure 10A, number
9), the DO concentration was higher at the second
monitoring port than at the first (Figure 11) because
of gas dispersion. DO concentration declined as the
air plug moved through the PFR.

Figure
11: ()

Operators also monitored pH in both the PFR and the STR (Figure 12). In that study,
the pH measurement along the PFR module did not indicate acid release. Monitoring
pH is especially useful when acids are formed in a PFR reactor for example,
in Escherichia coli cultivation.

Figure 12: ()

Looking Forward
The introduction of optical DO sensor
technology, intelligent sensors, and
wireless technology enables more reliable
biopharmaceutical
production
and
improved plant performance. Biomanufacturers will face challenges as they strive for
continuous improvement. In sensor technology, industry experts such as Lam would like
to see more measurement innovations, such as a reliable method for measuring
glucose online.
From a broader perspective, Leveen considers single-use technology to be the next
major trend in biopharmaceutical manufacturing. An early adopter of disposables for
biopharmaceutical production, he says, Because FDA regulation of therapeutics begins
with clinical trial material, the introduction of single-use bioprocess technology will begin
at this stage in therapeutic development.
Both industry suppliers and pharmaceutical company experts concur. Lam and others
insist that cost will be a driving factor in the decision-making process for years to come.
To succeed in the future biomanufacturing market, newly introduced technologies must
cost no more than current technology.
Acknowledgments
The authors are grateful to Stefan Junne, Arne Klingner, Dirk Itzeck, and Peter
Neubauer of the bioprocess engineering laboratory in the department of biotechnology
at the Technische Universitt Berlin for contributed content from the novel plug-flow
reactor case study as well as data and photos for Figures 10, 11, and 12.
The authors thank the following industry experts for helpful conversations: Dr. L. Harry
Lam, PhD, is a bioprocess engineer who has spent his career in biochemical

manufacturing operations for major pharmaceutical corporations. He has been

responsible for technology implementation in many successful development projects
and manufacturing campaigns. In his current role, he oversees outsourced contract
biological drug substance manufacturing for a major biopharmaceutical company.
Lindsay Leveen is a manufacturing process industry strategist and early technology
adoptor focused on sustainable development. He has led manufacturing technology
implementation for biopharmaceuticals, where he was an early proponent of single-use
methods. More broadly, he has implemented sustainable development for major
corporations in areas such as fuel cells, telecommunications, and power generation and
transmission. Currently, he is a consultant and recipient of the American Institute of
Chemical Engineers Professional Development award for his lifetime of achievement in
chemical engineering.
About the Author

Author
Details
Corresponding author Amber Ratcliff is market segment manager for analytical
sensors at Hamilton Company, 4970 Energy Way, Reno, NV 89502; 1-800-648-5950. A
consultant with GeneCom Group, Carol Preisig, PhD, MBA, provides scientific writing
and industry research assistance to the company. Arc, Polilyte, and Arc VisiFerm are
registered trademarks of Hamilton Company.

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