Tecan Journal
Edition 2/2018
Life Sciences, Diagnostics and Partnering
Building blocks for custom solutions
Redesigning the drug safety workflow
Cell counting made easy
Long-term partners in success
Pages 8-9
Pages 10-11
Pages 16-17
Pages 28-29
CEO WELCOME
Welcome Dear Reader, Automation is increasingly being seen as a ‘must have’ tool in the laboratory, not only because of the higher throughput it offers and the staff time it frees up, but also because of the improvements in precision and reproducibility it represents. This is especially true for regulated labs, where the shift in approach means that more and more laboratories with little or no previous experience using automation are choosing these technologies to accelerate and simplify their workflows. Automation is a particular strength of Tecan’s; through Tecan Partnering we supply more than 35 different diagnostic instrument platforms to more than 30 customers under their own labels, from leading diagnostics companies to segment specialists and emerging players. You can read more about companies following this trend in the Synchron (pages 8-9) and Vircell (pages 28-29) articles in this issue. Tecan Partnering has a broad and growing portfolio of OEM liquid handling options for the IVD industry, now including the exciting new Cavro® Magni Flex*, a modular, scalable, multichannel XYZ technology platform packed with features to make it easier for instrument designers to cost-effectively integrate advanced functions into their systems. We’re also taking a fresh approach to software development, with the introduction of MAPlinx™*, our modular application platform for OEM customers. This development framework combines standardized modules for functionality common to multiple systems with the ability to create custom application software accelerating development and time to market. We’ve news too of the end-user business where, following the introduction of the Fluent® Gx for regulated markets in the first regions in February 2018, we also registered the platform with the US FDA as a Class 1 Medical Device in April. Since the launch of the Fluent Gx, we have seen great demand for the newest liquid handling platform on the market, with some customers ordering several systems at once. Watch out for details in future editions of the journal. I hope you enjoy the issue, Dr David Martyr CEO
*Product coming soon. **MAPlinx is currently available for internal development only; for external development coming soon.
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TECAN JOURNAL 2/2018
CONTENTS
Contents 2
CEO Welcome
4 - 5 Putting stem cells at the heart of predictive drug toxicity testing
6 - 7 Is NGS costing you more than you think?
8-9 Building blocks for custom solutions
8 - 9
Building blocks for custom solutions
10 - 11
Redesigning the drug safety workflow
12 - 13
Sequencing made easy
14 - 15 Automation by numbers 16 - 17 Cell counting made easy 18 - 19
Long-read sequencing for improved analysis
20 - 21 Lipidomics – the future of personalized medicine? 22 - 23
Biosensors light the way for drug development
24 - 25 A tailored solution to meet bioprocessing challenges
28 - 29 Long-term partners in success
26 - 27 Making big discoveries accessible to all laboratories
28 - 29 Long-term partners in success 30 - 31 Detection at the molecular level
Applications and platforms presented in the Tecan Journal may not be available in all markets. Please consult your local Tecan office for information.
TECAN JOURNAL 2/2018
3
DRUG DISCOVERY
Putting stem cells at the heart of predictive drug toxicity testing Failure of candidate drugs in late-stage clinical development continues to be a challenging and costly problem for the pharmaceutical industry. A recent study of 640 novel therapeutics found that more than half (54 %) failed in clinical development, most (57 %) due to insufficient efficacy, and a sizable proportion (17 %) due to safety concerns.1 The earlier in the drug development process that these issues can be accurately assessed, the lower the risks – both financially and to human health – with taking a compound forward into clinical testing.
Stem cells are playing a growing role
electrophysiological changes that can
The protocols required to differentiate
in all phases of drug discovery – from
adversely affect myocardial contractility.
stem cells into the intended cell type
disease modeling to the development
In a comprehensive review article,
are equally complex, requiring close
of innovative therapies – providing
authors from the US Food and Drug
monitoring and the addition of precise
self-renewing, sustainable sources of
Administration discuss the advantages
amounts of various growth factors and
differentiated cell models. Derived
of using hiPSC-derived cardiomyocytes
other biological and chemical stimulants
from either human embryonic stem
to assess drug-induced structural
at key time points. Any inconsistency in
cells (hESCs) or induced pluripotent
cardiotoxicity.5 This article emphasizes
how these protocols are carried out – eg.
stem cells (hiPSCs), these models are
the renewable and readily scalable nature
due to variability between the technique
becoming increasingly popular for in
of stem cells, and the benefits of testing
of technicians – can affect the behavior
vitro preclinical predictive toxicology
directly in human cardiomyocytes rather
of the resulting cardiomyocytes in
testing, providing more physiologically
than having to translate findings from
subsequent toxicity testing.
relevant data to detect potentially
toxicity studies conducted in animal
harmful on- and off-target effects
models or animal cell lines. It concludes
Automated liquid handling and cell
before drug candidates reach the
that high throughput screening using
processing technologies can minimize
clinical phase.2,3 This helps to reduce
a hiPSC-cardiomyocyte model is a
this variability, as well as eliminating
attrition rates, costly program failures
‘feasible approach to assess potential
much of the tedious, time-consuming,
and post-market withdrawals, and can
contractile and structural cardiotoxicity
and labor- and cost-intensive aspects
even provide the opportunity for cost-
in early phase drug development’, and
of working with stem cells and stem
efficient modification of the drug to
supports combining structural and
cell-derived models. These benefits
minimize its toxicity.
electrophysiological tests on the same
become even greater as methods are
platform.
scaled up to meet the demands of large-
Using stem cells to detect drug
scale applications. High throughput
cardiotoxicity earlier
The challenges of working with
automation solutions are essential in
Cardiotoxicity is one of the top
stem cells
enabling stem cell-derived models, such
three causes of post-marketing
Stem cells are highly sensitive to
as hiPSC-derived cardiomyocytes, to be
drug withdrawals due to adverse
variability in their environment and
applied as a pharma industry standard
reactions. Rapid advances are being
handling, and can be tedious and tricky
for earlier predictive toxicity testing in
made in the use of hiPSC-derived
to work with. Nurturing stem cells to
drug discovery.
cardiomyocytes to test drug candidates
grow and reproduce in culture is as
for their potential to cause structural or
much art as science.
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The Blog 4
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 2/2018
DRUG DISCOVERY
References 1) Hwang, TJ et al. Failure of investigational drugs in latestage clinical development and publication of trial results. JAMA Intern Med, 2016, 176(12), 1826-1833. 2) MacDonald, A. Stem cells in drug discovery. Technology Networks, April 4, 2017. https:// www.technologynetworks.com/cellscience/articles/stem-cells-in-drugdiscovery-286825 3) iPS Cell Drug Discovery Taking Off with First Clinical Trial. Press release. August 2, 2017. https://asia. nikkei.com/Tech-Science/Science/iPScell-drug-discovery-taking-off-withfirst-clinical-trial
If you haven’t listened to it already, check out the recent webinar entitled Automated assessment of liver and cardiac toxicities in lead optimization, using biochemical and human iPS cell assays on the Tecan FluentŽ at www.drugtargetreview.com/ webinar/30994/automated-assessmentof-liver-and-cardiac-toxicities-in-leadoptimisation-using-biochemical-andhuman-ips-cell-assays/
4) Onakpoya, IJ et al. Post-marketing withdrawal of 462 medicinal products because of adverse drug reactions: a systematic review of the world literature. BMC Med, 2016, 14, 10. 5) Yan, X and Papoian, T. Moving beyond the comprehensive in vitro proarrhythmia assay: use of humaninduced pluripotent stem cellderived cardiomyocytes to assess contractile effects associated with drug-induced structural cardiotoxicity. J Appl Toxicol, 2018, Feb 27. doi: 10.1002/jat.3611. (Epub ahead of print)
www.tecan.com/blog TECAN JOURNAL 2/2018
5
GENOMICS
Is NGS costing you more than you think? The sequencing, analysis and data storage costs associated with NGS are dropping rapidly, making this powerful technology ever-more affordable and widely accessible. Cost analyses indicate that the landmark $100 genome is almost within reach, but the reality is that most NGS labs are still spending far more than that. Here we explore some hidden costs of NGS that occur upstream of sequencing, highlighting some of the ways labs are hemorrhaging money unnecessarily, without even realizing it.
Looking upstream of sequencing
Nucleic acid extraction – The ‘massively
or add something twice. At best, such
There have recently been several
parallel’ high throughput nature of
mistakes can mean that the process
articles looking at the ‘real cost of NGS’,
NGS inevitably highlights any upstream
needs to be repeated. At worst, errors
but most of these focus on everything
bottlenecks in sample preparation, often
such as cross-contamination can go
from sequencing onwards, and gloss
in nucleic acid extraction. Conventional
undetected, leading to erroneous results
over the upstream steps of sample and
DNA and RNA isolation methods are
and potentially serious consequences,
library preparation. Unfortunately, that
both low throughput and very labor
especially in a clinical setting.
is a significant oversight, as the NGS
intensive, decreasing productivity.
process starts long before you press
Manual extraction approaches also
PCR for library preparation and
‘go’ on your sequencer. The good news
increase the risk of inaccuracy and
quantitation – It is worth paying special
is that many of these ‘hidden’ costs can
human error, which ultimately translate
attention to PCR workflows that are
be easily reduced and controlled, if you
into wasted time, reagents and
integral to library preparation and
know where to look. The main culprits
resources. And, once extracted, nucleic
quantitation, as cross-contamination
in NGS are:
acids need to be accurately quantified
can be a major issue that is hard to
for subsequent titration. Using robotics
detect. The nature of PCR means that
to work with the smallest volumes
small errors are readily amplified and
misplace or mix up samples in a high
possible at this stage can significantly
can skew your library. For example,
throughput NGS workflow, leading
lower the price per sample.
pipetting the wrong index into the
Tracking sampling errors – It is easy to
wrong sample could lead to hugely
to re-collection or repeat testing. In some cases, replacing samples may
Library preparation – If you have
significant knock-on consequences.
not even be an option, or a mix-up may
large numbers of samples to process,
This can be particularly problematic
go undetected. Occasional errors like
library preparation can actually cost
when the aim is to assess rare
this may not seem like a big deal, but
more than the sequencing itself. With
variants (eg. when examining tumor
tracking down lost samples and re-
many steps involved for amplification,
heterogeneity). And, when PCR is used
running tests is a drain on resources,
pooling, normalization, etc., there are
for quantitation, everything is typically
and can add up to serious money. In
numerous opportunities for waste and
done in triplicate, multiplying the total
addition to the direct costs, there is
error. It is critical that reagents, samples
number of pipetting steps and different
also the incalculable impact on clients
and controls are added in the right
samples you have to deal with, further
and patients, ranging from simple
amounts, to the right wells, at the right
increasing the risk of errors. It is difficult
dissatisfaction to poor medical decisions
time. Since the volumes involved are
due to lack of information or an
very small – only a few microliters – it
erroneous result. End-to-end traceability
is easy to accidentally skip a sample
is therefore vital at every step of the NGS workflow, from sample collection right through to reporting.
The Blog 6
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 2/2018
GENOMICS
to estimate the frequency and ultimate
Check out our dedicated genomics
cost of these sorts of mistakes, as many
channel – The Helicase – at
of them may go undetected, so it is
www.tecan.com/helicase for
better to take measures to avoid them in
additional information and support on
the first place.
automating your genomics workflows.
Understanding the real cost of NGS This list is far from exhaustive, but hopefully the issues discussed here make it clear that taking time to scrutinize the NGS process from start to finish – especially the steps upstream of sequencing – is a worthwhile endeavor. Investing in quality automation can go a long way to addressing all of the above issues by eliminating unnecessary manual steps and sources of error. At the same time, it can free up skilled staff for more important (and interesting) work.
www.tecan.com/blog TECAN JOURNAL 2/2018
7
OEM SOLUTIONS
Building blocks for custom solutions Developing customized laboratory automation solutions for complex liquid handling operations requires access to state-of-the-art robotics and OEM instrumentation. Flexible and modular components are essential building blocks of many of these bespoke designs, and the Xantus® XYZ robot has stood the test of time, supporting over 10 years of OEM integration by Synchron Lab Automation.
Synchron Lab Automation – based in
company became a Tecan distributor
“Customers turn to us when a standard
Etten-Leur, the Netherlands – designs
around 1980, and my father – who also
pipetting platform does not meet their
and builds software and hardware for
worked in the business – was
requirements. Every client has an idea of
automated liquid handling systems,
subsequently asked to develop some
what they want, but we help to make it a
customized to its clients’ specifications
ELISA software for the Tecan MiniPrep,
reality, developing software and
and needs. The company has a long-
and this was the beginning of Synchron.
hardware and rapidly 3D printing
standing history of working with Tecan,
When Tecan decided to invest in direct
prototypes, prior to creating the final
as Derk Wilten, Director and Owner,
sales to its European customers, we
design. Our expertise in custom lab
explained: “Synchron was born out of
partnered with Sias, and now the story
automation has led to us working with
the automation department of Wilten
has come full circle; my father worked
customers in Europe, Singapore and the
Instruments, a laboratory supplier
with Tecan, then Sias, and now Sias is
USA, on projects ranging from benchtop
started by my grandfather in 1952. The
Tecan again.”
automation to more complex systems,
Derk Wilten, Director of Synchron Lab Automation
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TECAN JOURNAL 2/2018
OEM SOLUTIONS
The Xantus robot’s open geometry, modularity and flexibility have made it the best solution on the market for integration into our projects.
The Xantus robot offers a wide range of integration options
all dependent on our specialized
channels, with or without a gripper –
need additional training, we know where
knowledge of liquid handling robotics.
and we can even mount the arm to a
to turn. Tecan is a great company to
Rather than limiting ourselves to a single
wall, removing the deck entirely.”
partner with, and there’s no doubt that
market, we recognize that our expertise
the Xantus robot will continue to be a
can be applied across the board. Our
“The Flexispan feature – enabling
central feature in many of our future
customers therefore come from a full
individual channels to move
projects,” Derk concluded.
range of industries, including life
independently along the Y- and Z-axes
sciences, petrochemistry, pharma, food
– is also highly valued, and each channel
and beverage, and agriculture.”
can be fitted to a range of pumps and use either disposable or fixed tips. The
To find out more about partnering with Tecan, visit
“Although every project is different, they
new micropump technology also gives
all share some similarities, with liquid
us a competitive advantage, by allowing
handling at the heart of each design. If
us to accurately dispense both large and
we develop a technique to pipette milk
small volumes using the same pump.
To learn more about Synchron Lab
for a dairy, for example, the same
Our customers expect reliability and the
Automation, go to
technology can serve a similar need in a
system needs to be the best of the best.
www.synchronlab.com
blood bank. We look to build as much
The Xantus robot is extremely high
standardization as possible into our
quality and reliable. The precision
designs, and so it is essential to have
mechanics in the robotic arm are a feast
access to as many ‘building blocks’ as
for the eyes but, more importantly, they
possible, which is where the Xantus
are very easy to access for servicing –
robot comes into play. Its open
it’s a fine piece of Swiss engineering.”
partnering.tecan.com
geometry, modularity and flexibility have made it the best solution on the market
“We have now been working with the
for integration into our projects; 95 % of
Xantus robot for over 10 years, so we
our projects make use of this platform.
know exactly what configuration to buy
The configuration of the system is
and what to do with it. However, we still
amazing. We can get it in any size up to
meet with the Tecan team twice a year
three and a half meters long – with one
to discuss our projects and, if we ever
or two arms, up to eight pipetting
TECAN JOURNAL 2/2018
9
DRUG DISCOVERY
Redesigning the drug safety workflow Detecting adverse off-target
Safety pharmacology is an important
a-chip approaches – that enable drug
effects is crucial to ensure the
part of the drug discovery process,
safety studies to be performed far earlier
safety of potential therapeutics,
identifying and investigating any
in the development process. Ncardia is
potential undesirable pharmacological
a pioneer in this area, developing and
effects of candidate drugs before they
commercializing highly predictive cellular
enter clinical development. These effects
assay systems derived from human stem
have traditionally forced
are often hard to characterize in cellular
cells to accurately assess drug safety.
pharmaceutical companies to
or simple in vivo models, requiring
Dr Stefan Braam, Co-founder and CEO
perform safety pharmacology
animal testing – usually in canine models
of Ncardia, explained: “Our aim is to
studies at a late stage of the
– to ensure the potential therapeutic
put ‘human’ testing at the forefront
is safe to proceed to the next phase.
of drug discovery, by developing the
The cost and ethical issues associated
best possible human cellular models.
with such testing mean that safety
This is very attractive from an industry
models and automated screening
pharmacology studies have traditionally
perspective, as it ensures only viable
processes are revolutionizing
been performed at a late stage of
candidate drugs are progressed through
drug safety studies, enabling
preclinical development. This can lead
the pipeline, while avoiding the risks
much earlier testing, with
to late-stage failure of candidate drugs,
and ethical considerations associated
resulting in significant financial losses.
with animal testing. Our expertise in the
but limited throughput and ethical considerations
drug development process. Human stem cell-based cellular
companies such as Ncardia at
manipulation and differentiation of stem
the forefront of this workflow
Avoiding these expensive failures
cells into cardiac cell lines allows us to
transformation.
is a major goal for pharmaceutical
create a synchronously beating layer
companies, creating a real need for
of human cardiomyocytes in culture. In
new technologies – such as organ-on-
combination with predictive safety tests
Stefan Braam (left) and Farbod Famili with Ncardia’s Fluent Automation Workstation
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TECAN JOURNAL 2/2018
DRUG DISCOVERY
– such as multi-electrode arrays – these
applications. We therefore wanted
you the option to define everything –
cells provide a convenient and cost-
to automate the entire cell culturing
labware, protocol availability, dispense
effective cardiac model, allowing safety
process, performing each step as quickly
volumes, etc. – for inexperienced
studies to be performed much earlier
as possible to avoid variations across
operators, which makes it much easier
in the overall process. Our technology
a microplate. We looked at the various
for everyone in the lab to use the system
has already been adopted by many
liquid handling solutions on the market,
once it is in routine use. Working in
of the world’s top 20 pharmaceutical
and were impressed by both the speed
384-well microplates, we’re currently
companies, demonstrating the huge
and flexibility of the Fluent® Automation
handling 40 plates in a single run. This
potential of this approach.”
Workstation. With three independent
would take at least five times as long
arms, it offered us the potential to
to do manually, so the benefits are
Assay development is at the forefront
significantly accelerate the process, while
felt immediately.”
of Ncardia’s offering, and the company
still being a very versatile platform. We
is actively involved in the design and
had already had a positive experience
validation of assays intended for
working with Tecan when we purchased
day-to-day use by pharmaceutical
a Spark® microplate reader, and so we
and biotechnology companies. Stefan
were confident in both the service and
continued: “Safety pharmacology is just
support the company offers.”
one potential application of human-
For more information on Tecan’s drug discovery portfolio, visit www.tecan.com/drugdiscovery
derived cellular models, and we are also
“The system was installed at the
interested in using this technology to
beginning of summer 2017, and has
To learn more about Ncardia,
enable high throughput drug efficacy
everything required to culture and
go to www.ncardia.com
testing. By culturing cardiomyocytes in
differentiate stem cells integrated into
a 384-well microplate format, we can
it, including an automation-friendly
provide our customers with the tools to
incubator and a laminar flow HEPA
screen tens of thousands of potential
hood to ensure sterility. We knew that
drug molecules in a human model. This
setting up such a complex workflow
ensures more reliable and accurate
for highly sensitive cells would be
screening data, reducing the risk of
challenging, and finding the optimal
throwing out the baby with the
labware and workdeck configuration was
bath water.”
the first obstacle, but the 3D simulator in FluentControl™ makes it very easy
Farbod Famili, Assay Development
to visualize your protocols, speeding
Scientist at Ncardia’s Leiden facility
up process development. It also gives
in the Netherlands, took up the story: “Pharmaceutical companies understandably demand assays that are robust and reliable, and compatible with high throughput automated screening platforms. When I first arrived at Ncardia, almost all of our processes were performed manually, which worked when handling a limited number of compounds for safety pharmacology
We had already had a positive experience working with Tecan… so we were confident in both the service and support the company offers.
studies using multi-well, multi-electrode array chips, but was not compatible with high throughput efficacy testing
TECAN JOURNAL 2/2018
11
GENOMICS
Sequencing made easy Automation is helping the University Hospital Münster to streamline its molecular diagnostics workflows, increasing throughput for Sanger sequencing and NGS library preparation.
The University Hospital Münster (UKM)
workflows, as Dr Hartmut Schmidt, Head
Hartmut continued: “One of the major
is one of Germany’s leading hospitals,
of Molecular Diagnostics, explained:
challenges in Sanger sequencing is
providing state-of-the-art healthcare to
“One of our department’s main roles is
the removal of unincorporated dye
national and international patients. It is
to perform routine Sanger sequencing
terminators. In the past, we did this
also home to the Center for Laboratory
on samples from several different
manually, but it was a slow process that
Medicine, which provides core
groups. We can carry out as many as
required three hours of preparation
laboratory services – including clinical
2,000 analyses per day, occasionally
before sequencing could begin. Not
chemistry, hematology, endocrinology,
even more; the human genetics
only was this time consuming, but it
virology, immunology and therapeutic
department, for example, sometimes
restricted our flexibility in the lab, and so
drug monitoring – to the hospital and
submits nine plates for processing on
we turned to automation.”
its external clients, typically performing
one day, which is around 900 samples
around 10,000 analyses per day. The
from just one group.”
laboratory’s molecular diagnostics department uses Sanger sequencing with capillary electrophoresis for routine diagnostics, as well as performing next generation sequencing (NGS) in collaboration with other UKM groups. Automation is the key to standardizing and streamlining the department’s
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TECAN JOURNAL 2/2018
One of the big advantages of automating the workflow is the elimination of dye blobs… enabling faster analysis of the data.
GENOMICS
Left to right: Mathias Isakiewitsch, Hartmut Schmidt, Kathrin Meyer zu Himmern and Kathrin Haar with the Freedom EVO workstation
The laboratory chose a Freedom EVO®
to medium to high; if the signal is too
chemistries just by using different
150 workstation equipped with a
high, the analysis may fail and have to
automated methods; it’s a really good
MultiChannel Arm™ 96 to help with
be repeated. We use a magnetic bead-
system, and one that can evolve to
the dye removal. This set-up allows
based method of purification, and have
accommodate other chemistries in the
one plate to be incubated while the
been able to implement a signal cut-off
future as our needs change.”
workstation performs the necessary
value by using the Freedom EVO to
pipetting steps to prepare the next,
add a defined amount of beads. This
In a busy department, staff only have a
which is an important feature to meet
allows us to remove any unincorporated
limited amount of time to spend learning
the lab’s testing demands. “Automation
dye terminators, maintaining the signal
a new technique, and one of the big
of the clean-up procedure has improved
intensity within a suitable range and
advantages of the Tecan systems for
our throughput – we can do up to six
reducing the number of repeat sample
UKM is that they are easy to use. The
runs in one hour – allowing us to handle
analyses, something we were unable to
company’s service engineer helped UKM
a greater number of samples and report
do with our previous filtration-based
staff to write TouchTools™ protocols that
the results more quickly,” Hartmut
purification process.”
take operators through the workflows
commented.
step by step, using photographs integrated In addition to Sanger sequencing, the
into each program to display on screen
“As an accredited laboratory, we take
molecular diagnostics department
exactly where the labware, samples
great care to ensure the quality and
performs automated NGS library
and reagents should be placed on the
reliability of our sequencing results,
preparation for a number of other
workdeck. “It is a really useful feature that
and thoroughly compared our old
UKM groups. This work is performed
makes the platform very straightforward to
method with the automated protocol
on a Freedom EVO NGS workstation,
use,” Hartmut concluded.
before it was implemented. One of
which is available with a number of
the big advantages of automating
verified protocols – developed in
the workflow is the elimination of dye
collaboration with the vendors – for
To find out more about Tecan’s
blobs – artefacts that interfere with the
different chemistries. Tecan application
genomics solutions, visit
sequencing analysis, making it harder to
specialists worked with the UKM team
www.tecan.com/genomics
identify the correct base pairs – enabling
to implement and evaluate these
faster analysis of the data. We have
protocols for the chemistries currently
also been able to overcome the issue
used by the human genetics and
of variations in signal strength. The
neurogenetics departments. Hartmut
samples sent to us for analysis by other
added: “Now that the evaluation is
groups vary in signal intensity from low
complete, we can process these varying
To learn more about UKM’s Center for Laboratory Medicine, go to www.ukm.de
TECAN JOURNAL 2/2018
13
BIOPHARMA
Automation by numbers Biopharmaceutical production is a complex process compared to small molecule drug manufacture, requiring the interaction of multiple biosynthesis pathways to create the target product. KBI Biopharma is using high throughput bioanalysis to gain an in-depth understanding of these biological processes, ensuring efficient, reproducible and scalable biomanufacturing processes. Carolina, and works with a wide range of
analyst – which we were able to achieve
clients – from biotech start-ups to big
and validate within six weeks of
pharma – to accelerate and optimize
installation. This automated version of
their drug development. The company’s
the assay offered at least comparable
highly successful biological
performance to our manual assay, and
manufacturing programs rely on using
took the same amount of time to run
advanced analytical techniques to
– 3.5 hours. We then looked at how to
acquire in-depth knowledge of each
use the instrument’s worktable capacity
biosynthesis pathway, resulting in more
and scheduling capabilities to save time
robust and scalable processes.
by processing multiple assay plates in parallel, something that would be
This data-driven approach requires large
virtually impossible for a single analyst
numbers of samples to be tested for
to achieve manually.”
each new project, and the company’s analytical development team recently
Devon continued: “The first step for any
Members of the analytical development team
began investing in automated liquid
parallel assay is to work out the on-deck
(left to right): Kirsten Dysinger, Elijah Gaspard,
handling solutions to enhance its high
capacity of every element required to run
Devon Davenport, Phoebe Lee and Derek Ryan
throughput testing capabilities. Devon
the assay – including hardware modules,
Davenport, Scientist I, explained: “When I
assay plates, labware, consumables and
arrived at KBI in 2014, there was no
reagents – then back-calculate the
automation in use for our ‘platform’
maximum throughput based on the
assays; everything was performed
limiting factor. For the HCP assay, the
Biopharmaceutical production requires a
manually. Coming from a high
volume of the reagent trough determined
wide range of studies – including
throughput automation background, I
that we could run eight plates without
immunoassays, enzymatic assays,
could immediately see the benefits of
the need for user intervention, but there
residual DNA detection and
moving some of our high volume residual
were only six assay positions on the
chromatography – to fully understand
assays to an automated platform, and
worktable. We therefore built a script
and optimize the biomanufacturing
started to explore the options available.
around parallel processing of six assay
process. Many of these analytical
My manager, Alan, and I each already had
plates. This protocol took just under five
activities are both time consuming and
some experience with Tecan platforms
hours to run all six plates – a huge time
complex, leading many
from previous roles and so, in July 2016,
saving – but still resulted in instrument
biopharmaceutical manufacturing
KBI purchased a Freedom EVO® 200 to
‘idle time’ and unused reagent capacity,
facilities to use automated or semi-
automate our HCP ELISAs. This one
and so we ‘looped’ two additional assay
automated workflows. This not only helps
assay was occupying four members of
plates onto the end of the protocol by
to increase throughput and improve
staff full time, and our intention was to
exchanging the completed plates with
consistency, it also frees up technicians
offload all of this work onto the
two fresh ones from the instrument’s
to perform tasks that are less automation
automated platform, allowing the
labware hotel. As a result, we can now
friendly or to analyze results.
analysts to perform other activities.”
process eight complete assay plates in 9
KBI Biopharma is a contract development
“We began by setting up a ‘single plate’
for user intervention, compared to 28
and manufacturing organization
assay protocol on the Freedom EVO –
hours of hands-on analyst time. We have
headquartered in Durham, North
effectively taking the place of one
since applied this same approach to a
hours and 37 minutes without the need
14
TECAN JOURNAL 2/2018
BIOPHARMA
The systems’ Freedom EVOware Plus has been great to work with; it’s very user friendly and intuitive.
number of other HCP ELISA protocols – each designed to meet individual clients’ specifications – with similar results.” “Following the success of the HCP project, we have now also begun automating our residual DNA platform method using a Freedom EVO 100, based on the positive experience we’d had with the HCP platform and the ease of scripting within the software. The systems’ Freedom EVOware® Plus has been great to work with; it’s very user friendly and intuitive, and offers advanced users the ability to change and modify far more parameters than most other laboratory automation software I’ve worked with. This allows you to edit, for example, the exact timings and durations of individual pipetting steps or labware movements, which proved critical to achieving an optimized six-plate HCP assay. Building on this, we have now created modular automation ‘nodes’ which we can link together to form a complete protocol using the different, client-specific HCP assays running in parallel. The local Tecan team – who happen to be just up the road – have been great to work with, and have been extremely supportive as I’ve tried to get the most out of the system. Introducing the Tecan platforms has been so successful that we’re considering an
To find out more about Tecan’s
additional Freedom EVO 200 to provide
Freedom EVO platform, visit
extra capacity and allow more assay development,” Devon concluded.
www.tecan.com/freedomevo To learn more about KBI Biopharma, go to www.kbibiopharma.com
TECAN JOURNAL 2/2018
15
CELL IMAGING
Cell counting made easy Small molecule drug discovery involves a range of functional assays that have traditionally relied on manual cell counting techniques to monitor proliferation, migration and invasion. Automated cell counting is enabling the EB House Austria to save time and free up personnel, as well as designing time-course experiments that were previously unachievable.
The EB House Austria is a unique,
while the gene therapy team is hoping
entered phase III clinical trials. Our
highly specialized clinic for patients
to tackle the problem at its root, by
attention then turned towards treating
with epidermolysis bullosa (EB), a rare
replacing or correcting genes using
RDEB, where we are currently studying
inherited disease that causes the skin to
different technologies, such as
differently regulated microRNAs in
become ‘as fragile as a butterfly’s
CRISPR, conventional DNA therapy or
developing tumors to identify potential
wings’, blistering in response to friction
trans-splicing.”
biomarkers.”
or trauma. The clinic, located within the University Hospital of Salzburg, was
“Our research covers the full spectrum
“A large variety of functional assays
the initiative of patient organization
of the disease, from the milder EB
were needed to support our research,
DEBRA Austria, and opened in 2005.
simplex (EBS) to the most severe form
and we purchased a Spark® multimode
The EB House consists of three units:
– recessive dystrophic epidermolysis
microplate reader back in 2016 to meet
the EB outpatient clinic; the EB
bullosa (RDEB) – which ultimately leads
the workload. We carry out confluence
academy; and a research arm
to premature death from chronic
measurements for many proliferation,
developing therapies to treat the
wounds that develop into cancers.
migration and invasion assays,
disease. The clinic’s 30-strong group
Our initial focus was on EBS, where
as well as using luminescence to
of researchers is divided across three
we discovered an inflammatory pathway
study the interactions of microRNAs
areas: gene therapy; basic research into
that is constitutively activated in the
with three prime untranslated regions
cancer and wound healing; and small
patient population. We repurposed the
(3’-UTRs), and to assess cell viability
molecule development and epigenetics.
interleukin-1 beta inhibitor diacerein –
with tetrazolium dye MTT assays.
Dr Verena Wally, Group Leader of
which is used in the treatment of
We also investigate transfection
Small Molecules Research, explained:
osteoarthritis – to treat these patients.
efficiencies, for example by
“The main aim of my group is to
This project has since been out-licensed
looking for green fluorescent
identify potential therapeutic targets,
to a pharma company, and has now
protein expression.”
Members of the EB House research team (left to right): Verena Wally, Michael Ablinger, Thomas Lettner, Roland Zauner, Monika Wimmer and Melanie Böhm
16
TECAN JOURNAL 2/2018
CELL IMAGING
eproducibility R has been a key advantage of the new platform, as the reader removes any user bias or human error, and we can now measure even smaller cell sizes.
“In September 2017, we purchased the
scale migration assays involving 12
“We chose to purchase the Spark on the
Gas Control Module (GCM™) and Live
plates, something we couldn’t do
back of a conversation with another lab,
Viewer, which offered us the expanded
previously. Ultimately, the greatest
and Tecan’s reputation for excellent
functionality we need. The reader has
benefits have been the time saved and
customer service. We have been
brought huge benefits to our workflow.
the personnel it frees up. The platform
impressed by the responsiveness and
Previously, we were carrying out
now runs every day, and is used by all
level of technical support that we have
migration assays by hand; taking
the groups.”
received. We’re fortunate to be only 20 kilometers from the Tecan office and, if
pictures with a microscope and using cell counting software or counting them
“We are currently preparing to run
we have a problem, a Tecan team
ourselves. For luminescence assays, we
time-course experiments over 24 hours,
member gives us a call or visits the site;
were fortunate to be able to use
measuring every two hours to monitor
it was a deciding factor in choosing to
systems from other departments, but it
the proliferation and viability among
upgrade the system,” Verena concluded.
was time consuming to move between
other factors. Before the Spark,
locations, particularly when running
we were limited to taking
time-course experiments.
measurements of time points when
Reproducibility has been a key
we were actually in the lab – say after
To find out more about live cell
advantage of the new platform, as the
eight hours, 12 hours and 24 hours –
imaging, visit
reader removes any user bias or human
but this could lead to us missing
error, and we can now measure even
the time point that would actually be
smaller cell sizes, expanding the range
important. Leaving the plate in the
of cell types we can analyze. In
reader and taking readings every two
addition, the Spark can handle higher
hours will be hugely beneficial. Gone
density plates, enabling us to measure a
are the days when if we missed the
House Austria, go to
greater number of samples in a single
event, we had to start again from
www.eb-haus.org/en/home.html
run. We can also now carry out large-
the beginning.”
lifesciences.tecan.com/live-cellimaging To learn more about the EB
TECAN JOURNAL 2/2018
17
GENOMICS
Long-read sequencing for improved analysis Next generation sequencing is now in widespread use throughout the life sciences sector, but the commonly used short-read sequencing methods are often subject to GC base pair bias. Combined with the inherent mapping ambiguity of the short reads, this often results in fragmented genome assemblies, creating a demand for technologies offering longer reads that simplify analysis and yield more complete sequences. Using its proprietary technology, Pacific Biosciences is able to offer longer reads, more uniform coverage and high accuracy, supporting advanced genomics, full-length transcript sequencing and epigenetics.
“The major advantages of our technology are the long reads, uniform coverage and high consensus accuracy. This accuracy is due to low sequencing-context bias, very low systemic error and the random nature of read errors. For instance, with 10- to 20-fold coverage we can achieve 99.9 % accurate reads, which can be readily increased to 99.999 % accuracy with additional sequencing coverage. Our system can also accommodate a broad range of template sizes – from ~200 to >10,000 bp – making it suitable for a wide variety of applications, including whole genome and targeted sequencing. As a result, SMRT Sequencing technology is proving popular for plant and animal sciences research – particularly in academic labs – as well as in the human biomedical space, for applications
The Cavro ADP is at the heart of the Sequel System
such as HLA typing.” Pacific Biosciences (PacBio) is a
(ZMWs), which you can think of as very
sequencing technology provider based
small reaction vessels. We anchor the
“Our original system – the PacBio RS II –
in Menlo Park, California. Using its
polymerase with the pre-bound DNA
had 150,000 ZMWs per SMRT Cell,
proprietary Single Molecule, Real-Time
template at the bottom of the ZMW,
equating to about 55,000 reads, and
(SMRT®) Sequencing technology, the
and introduce fluorescently-tagged
could process up to 16 cells per run. This
company has developed two long-read
nucleotides. The ZMW is illuminated
may sound like a very large number, but
sequencing systems – the PacBio RS
from below, and the attenuated light
throughput was still a limitation, so we
II and the Sequel® System – offering
enters the lower 20-30 nm of each
launched the Sequel System in October
a comprehensive view of genomes,
ZMW, exciting the fluorophore as the
2015. With this platform, each SMRT Cell
transcriptomes and epigenomes.
polymerase incorporates each base
contains one million ZMWs, offering a
Marty Badgett, Senior Director of
into the native strand. Our instrument
sevenfold improvement in throughput,
Product Management at PacBio,
basically carries out signal processing
and we’re currently working on a higher
explained: “Our instruments sequence
by ‘eavesdropping’ on the process of
density SMRT Cell containing eight
DNA directly in SMRT Cells packed with
DNA synthesis as it happens.”
million ZMWs.”
thousands of zero-mode waveguides
18
TECAN JOURNAL 2/2018
GENOMICS
Automation of NGS protocols is now crucial for many life sciences applications
Paz Yogev, a Staff Mechanical Design Engineer at PacBio, added: “When we were developing the Sequel System, we wanted to incorporate a number of additional liquid handling features that the pipettor used in the PacBio RS II simply couldn’t offer. We researched a
his OEM solution offers a combination T of performance, accuracy, cost and advanced features… that is very favorable for our application.
number of off-the-shelf solutions – as well as the possibility of developing a customized pipetting system based on one of these options – before eventually deciding on Tecan’s Cavro
®
Air Displacement Pipettor (ADP). This OEM solution offers a combination of performance, accuracy, cost and advanced features – including automatic tip detection and ejection, plus liquid level detection – that is very favorable for our application. We initially bought a couple of sample units and set up a test rig to run comparisons. This was both easy and inexpensive, making development of the pipetting system
same supplier maximizes the likelihood of everything working in synergy
To find out more about
without any issues.”
partnering with Tecan, visit partnering.tecan.com
“We have been working with the Cavro ADP for several years now, and it has been really good to us – it’s very reliable. Our field service engineers respond to any problems encountered
To learn more about Pacific Biosciences, go to www.pacb.com
by customers, and we very rarely have to reach out to the Tecan team for extra support, but we know it’s there if we need it,” Paz concluded.
very cost effective. In addition, we chose to use Tecan’s disposable tips, as buying the pipettor and consumables from the
TECAN JOURNAL 2/2018
19
MOLECULAR DIAGNOSTICS
Lipidomics – the future of personalized medicine? Personalized medicine is on the horizon, and cell membrane lipidomics may hold the key. Italian biotechnology company Lipinutragen is studying the relationship between the membrane status and dietary and health conditions. A crucial part of the process is the isolation of mature red blood cells, which can be performed more precisely and reproducibly using laboratory automation than by manual processing. of a balanced diet. The main cause of a nutritional imbalance is ignorance. People know that a lack of fruit in the diet can lead to a vitamin deficiency, but are simply not aware of the need to consume omega-3 and -6 fatty acids, which can lead to the development of essential fatty acid deficiency syndrome.” “Sample preparation for lipidomic analysis is a laborious, time-consuming, multi-step process that is difficult to perform precisely by hand, and so we automated the workflow on a Freedom EVO® platform. Blood samples are collected in tubes containing EDTA – the only anticoagulant able to The Lipinutragen team (left to right): Andreea Popa, Carla Ferreri, Anna Vita Larocca, Simone Deplano, Valentina Sunda and Chryssostomos Chatgilialoglu
sequester calcium – and delivered to the laboratory within 24 hours. After centrifuging, plasma, platelets and lymphocytes are removed leaving the
Bologna-based Lipinutragen, a spin-out
and specialist pharmacies, which
red blood cell layer. Red blood cells have
of the Italian National Research Council
can offer advice about dietary
a four-month life span, and the mature
(CNR), was founded in 2005 by
changes and supplements to address
cells that we require are smaller in
Drs Chryssostomos Chatgilialoglu
a lipid imbalance.”
diameter and heavier than the others. We
and Carla Ferreri, who drew on their
measure the diameter of the red blood
expertise in synthetic and analytical
Carla continued: “Typically, the first
cells using a SCEPTER™ 2.0 handheld
chemistry to establish a lipidomics
sign of a lipid imbalance is stress.
cell counter (EMD Millipore), selecting
laboratory, focusing on the isolation
Biochemical analysis may not give
those that are six microns across.
of mature red blood cells to allow
any indication of a pathology, but
These cells are removed and placed in
analysis of the membrane fatty acid
the person’s mood may change, or
water to rupture the membrane, and
profile. Carla, now Lipinutragen’s R&D
they might not sleep well and still
then centrifuged to produce a pellet.
Director and a Senior Researcher at
feel tired when they wake up in the
The phospholipids are extracted from
CNR, explained: “The lipid profile of the
morning. These are all signs of a general
the pellet using chloroform:methanol
mature red blood cell membrane is a
imbalance, including the cell membrane
(2:1, v/v) and derivatized to form fatty
comprehensive biomarker of a person’s
lipid profile. The membrane contains
acid methyl esters for analysis by gas
state of health, quality of nutrition and
two types of fats; those that can be
chromatography. All stages except
lifestyle. This has benefits in both clinical
produced by the body’s enzyme system,
centrifugation, evaporation and GC
applications – enabling the development
and those such as omega-3 and -6 fatty
analysis have been automated, which has
of personalized treatment strategies –
acids, which must be consumed as part
made a big difference to our laboratory.”
20
TECAN JOURNAL 2/2018
MOLECULAR DIAGNOSTICS
The result was a liquid handling system that is far more consistent and precise than manual pipetting, which is exactly what we need for lipidomic analysis.
Automation of the process on the Freedom EVO has enhanced precision
“We chose the Freedom EVO because
or aspirate the extraction solvent, which
we needed a platform that could offer
is important for quantitation. To ensure
both precise tip movements and near
total confidence in our results, we run
perfect handling of a variety of solvents,
a monthly calibration procedure to
including water, chloroform, methanol
verify the automated extraction and
and hexane. The system also needed to
derivatization of a lecithin standard of
be able to accurately remove the mature
known concentration, quantifying the
red blood cell layer, and so we worked
results by GC analysis.”
with the Tecan Integration Group to
to be replaced with high throughput, automated systems, and so the next step is to gain high level accreditation from an international body for the protocol we have established. This will open the door to potential new projects at Lipinutragen and attract interest from other laboratories engaged in this type of work. Our aim is to make this platform available to other laboratories
customize the platform to mimic our
“Determining the membrane lipid profile
manual pipetting procedures, setting up
is the key to distinguishing between
and refining unique software to control
a pathological problem and essential
all the automated movements. The
fatty acid deficiency syndrome, and we
result was a liquid handling system that
are raising awareness of this concept
is far more consistent and precise than
across the medical field through training
To find out more about Tecan’s
manual pipetting, which is exactly what
courses registered with the Italian Health
Freedom EVO platform, visit
we need for lipidomic analysis.”
Ministry. These courses are attended
www.tecan.com/freedomevo
around the world, helping to standardize lipidomic analysis,” Carla concluded.
by doctors, pharmacists and biologists “Precision is essential from the outset,
working in the fields of nutrition and
To learn more about
starting with the isolation of the mature
metabolic problems, allowing them to
red blood cells. Removal of the various
extend their knowledge of the subject
Lipinutragen, go to
layers above these cells is a crucial step
for accreditation and continuing
and must be performed identically each
professional development purposes.
time, which was extremely difficult to do
I believe that our lipidomic analysis
manually. Reproducible liquid dispensing
protocol will become commonplace in
and aspiration was also challenging; with
the future, with people having an annual
automation, we can precisely dispense
assessment. To keep pace with this
the required volume of solvent or water,
demand, manual processes will need
www.lipinutragen.it/en
TECAN JOURNAL 2/2018
21
DRUG DISCOVERY
Biosensors light the way for drug development The successful treatment of
The Moulder Center for Drug Discovery
biosensors, I can identify compounds
inflammatory diseases may lie
Research in Philadelphia, USA, serves as
that activate certain antioxidant
with controlling the production
a multidisciplinary research hub for
pathways, which may have therapeutic
Temple University School of Pharmacy,
applications.”
of particular proteins, driving efforts to identify translational
one of the oldest schools in Pennsylvania. The center focuses its
Oscar continued: “The types of things
repressors for drug targeting.
efforts on addiction withdrawal,
we can now achieve in biological
Scientists at the Moulder Center
Alzheimer’s disease, cancer, anti-viral
research are increasingly complex. There
for Drug Discovery Research
agents, and cardiovascular and
are many tools to investigate different
have developed luminescence-
metabolic diseases. Dr Oscar Perez-Leal,
properties of cells, and you can use one
an Assistant Professor, explained: “My
device to study multiple things
research focuses on drug development
simultaneously. For example, it is
for inflammatory diseases, in particular,
possible to carry out multiplex studies
multiplex studies and time-
fatty liver disease – which is increasingly
involving both luminescence and
course assays for identifying
prevalent due to obesity – and psoriasis,
fluorescence, the latter being used to
and characterizing novel
both of which have a high degree of
monitor cell viability. This approach
compounds.
inflammation and related oxidative
provides much greater insight into how
stress. My primary interest is in
cells respond to drugs, but depends on
identifying translational repressors of
having an instrument that also offers live
proteins that could become new drug
cell incubation, as well as
targets. In the last few years, I have
monochromators to allow selection of
created luminescence-based biosensors
the appropriate wavelengths for
– basically living cells that will only emit
excitation and emission. The Spark®
light when, for example, the target
multimode microplate reader – which
protein is being produced – and have a
was purchased in December 2017 – is
couple of related patents. Using these
well suited to meet these requirements.”
based biosensors for protein detection, supporting
22
TECAN JOURNAL 2/2018
DRUG DISCOVERY
I t’s convenient to have a multifunctional platform to handle routine screenings, as well as more infrequent specialized applications, which generates high quality data every day of the week – it’s fantastic! Dr Perez-Leal
“In order to evaluate the capabilities of
incubation as well. I wanted to combine
For example, I can determine when a
the reader, I borrowed a system from
everything I might need in a single
compound reaches maximum activity or
Tecan for a week to test it out,
platform, allowing me to run lots of
the lowest concentration that shows a
supported by a company rep for the first
different assays, which I simply couldn’t
therapeutic effect, and it’s great to be
three days. I did a fair amount of
do previously. For example, developing
able to observe all of this in real time. It
preparation in advance, so I could spend
live cell systems involves DNA
makes such a difference being able to
the week setting up assays, using the
manipulation and molecular biology, so
leave the plate in the reader and not
system as much as possible and learning
quantifying DNA and protein
have to transfer it backwards and
how to operate it, as well as making
concentrations using the NanoQuant
forwards to an incubator. Before the
comparisons with other instruments
Plate™ is ideal. It’s convenient to have a
Spark, I simply didn’t have this
in terms of sensitivity and speed.
multifunctional platform to handle
functionality in the lab; it was impossible
I experimented with many of the Spark’s
routine screenings, as well as more
to do this with my previous set-up. I’m
parameters that week, and I really liked
infrequent specialized applications,
very satisfied with the instrument, and
the fact that I was able to use it and
which generates high quality data every
I’ve enjoyed working with the Tecan
understand how it works fairly quickly.
day of the week – it’s fantastic!”
team – they’ve been very supportive,”
As a result, when the system was
Oscar concluded.
purchased, I already had some basic
“The control module is intuitive and easy
assay protocols set up, and have been
to use, and it’s straightforward to select
able to get a lot of data in a short space
the steps you need. The software
To find out more about Tecan’s
of time.”
arranges them sequentially so I can see
Spark reader, visit
the overall workflow from top to bottom.
www.tecan.com/spark
“It’s not uncommon for labs to purchase
One of the great benefits of the Spark is
an instrument for a particular project
being able to identify compounds with
without thinking of the wider, long-term
interesting functionality – say at the
provision. They might only need a
12- or 24-hour time point – and then
platform with luminescence capabilities,
isolate them to run time-course assays
and then discover further down the line
using different concentrations, which
that they would have liked live cell
offer a wealth of information.
To learn more about the Moulder Center for Drug Discovery Research, go to moulder.temple.edu
TECAN JOURNAL 2/2018
23
BIOTECHNOLOGY
A tailored solution to meet bioprocessing challenges Automation offers many benefits for bioprocess development involving multiple microbial strains, yet few off-the-shelf platforms are capable of combining liquid handling tasks with microscale cultivation. The Microbial Bioprocess Lab at Forschungszentrum Jülich needed a customized platform to support its research, and turned to the Tecan Integration Group for a solution that has improved its microbial cultivation workflows.
Forschungszentrum Jülich in western
“The BioLector can run up to 48
is based just 30 km away. This allowed
Germany is one of the biggest research
experiments simultaneously in
in-depth discussion between all the
facilities in Europe, with around 6,000
microplates, which is far more productive
parties involved, ensuring that the TIG
employees. The government-funded
than working with a few bioreactors that
team had a thorough understanding
organization has multiple institutes on
require continual monitoring. However,
not only of MiBioLab’s needs, but
campus, tackling a diverse range of
we still faced the challenge of sampling
also of the BioLector and how this
fields from supercomputing and physics
and dosing, which often has to be done
microbioreactor system could be
to informatics and biotechnology. The
during the night or according to the
integrated into a liquid handling
Helmholtz Innovation Lab ‘Microbial
real-time process data. You don’t
platform. TIG’s biggest challenges were
Bioprocess Lab’ (MiBioLab) is part of
want to have somebody sitting in front
to develop a system that had sufficient
the Institute of Bio- and Geosciences,
of a bioreactor 24/7, so this motivated
space to accommodate the BioLector,
and works with industry to investigate
us to find an alternative solution.
and to coordinate movements between
the use of renewable resources for
Automation was clearly the answer,
the two platforms to enable precise
fast and cost-effective production
so we spoke to a number of providers,
sampling and dosing on a shaking
of sought-after compounds. Dr. Ing.
including Tecan.”
microplate. The result was a customized
Holger Morschett, Head of MiBioLab,
workstation with a novel configuration –
explained: “The focus of our research
The Tecan Integration Group (TIG)
using the workdeck of a Freedom EVO®
is the use of microbial cell factories
visited MiBioLab to see the BioLector
150 adjacent to the BioLector system,
to convert renewable resources
and talk through the laboratory’s
combined with the liquid handling
– for example, biomass-derived
requirements, and was joined by
and robotics capabilities of the larger
carbohydrates – into sustainable value-
representatives from m2p-labs, which
Freedom EVO 200.
added products. This could be anything from chemical intermediates for the production of bioplastics to amino acids, organic acids and drugs, as well as industrially- and pharmaceuticallyimportant proteins. Our main tasks are strain phenotyping and bioprocess development, and the large number of genetically-engineered microbial strains that we need to test is a constant challenge. It’s impossible to do this in standard bioreactors, so we have transferred our protocols to a smaller scale system – the BioLector® microbioreactor (m2p-labs) – to give us higher throughput and allow easy automation.” Left to right: Marco Oldiges, Carin Jansen and Holger Morschett with MiBioLab’s TIG system
24
TECAN JOURNAL 2/2018
BIOTECHNOLOGY
TIG developed a customized platform for MiBioLab
I ntegration of the microbioreactor with the Freedom EVO has allowed us to perform experiments in a completely different way, giving us the opportunity to develop and test processes that we simply couldn’t do before.
The clear benefits of this approach led to us purchasing a second platform in January 2018, and both systems are now in use 24/7; I often have to book several weeks ahead when I need to use one of them!” “Working with the system is straightforward. We had some training
Holger continued: “A deciding factor
do with the BioLector, and allowed us
in choosing Tecan was the company’s
to gather the type of data you might
ability to guarantee what no other
expect from a well-equipped lab-scale
provider could. The microplate is
bioreactor. For example, we can take
sealed with a composite foil to prevent
samples and measure substrate and
contamination, and is continuously
product concentrations around the clock
shaken in the BioLector to ensure that
– something we couldn’t normally do
the cells have a sufficient oxygen supply.
with microliter volumes.”
Previously, shaking had to be interrupted
on Freedom EVOware® when the first platform arrived, and had simple workflows in operation after just four weeks, and more complex ones within three months. We were able to get our second system up and running after just one week of testing and transferring scripts. We have welcomed the support from Tecan, particularly for software
for sample extraction or dosing, leading
“Harvesting cells at the right time
integration, and know where to get
to transient oxygen deficiency and
point is not particularly easy to
help if we need it. It’s all working well,”
problems with cell viability. Of all the
manage manually, but automation
Holger concluded.
suppliers we approached, only Tecan
has provided the solution, thanks to
was able to guarantee that its solution
two-way communication between
could pierce the foils and carry out the
the microbioreactor and Freedom
appropriate liquid handling tasks while
EVO system. The BioLector measures
the plate was still shaking at speeds of
parameters such as biomass, pH
up to 1,500 rpm.”
and dissolved oxygen levels in real time, then notifies the liquid handling
“Integration of the microbioreactor with
platform of the location of wells that
To find out more about the
the Freedom EVO has allowed us to
need to be sampled or harvested, and
Tecan Integration Group, visit
perform experiments in a completely
the aspiration volume required. The
www.tecan.com/tig
different way, giving us the opportunity
Freedom EVO has everything required
to develop and test processes that we
for sample processing integrated onto
simply couldn’t do before, using robotics
the workdeck – including a centrifuge,
to fully automate tasks such as media
vacuum filtration station, microplate
preparation, sampling, dosing and cell
shaker, microplate cooler and an
separation. The integrated platform
Infinite® M200 microplate reader – so
has expanded what it is possible to
samples can be processed immediately.
To learn more about MiBioLab and see the automated platform in action, go to www.fz-juelich.de/mibiolab
TECAN JOURNAL 2/2018
25
DRUG DISCOVERY
Making big discoveries accessible to all laboratories Developing therapies to treat
Acute myeloid leukemia (AML) is an
working on two main projects; one for
rare diseases is often hindered
aggressive cancer affecting fewer
AML and the other for T-cell lymphoid
by the limited availability
than 10 in 100,000 people each year
leukemia (TLL). The AML project
in Europe. The resulting scarcity of
started as a chemogenomics screen
primary patient samples for in vitro
for a modulator in primary AML, and
testing presents a challenge for drug
we are now validating the result of this
samples, it is difficult to
discovery laboratories seeking out novel
using the various models available to
understand the fundamental
compounds to treat them. A recently
us in the lab. The second project stems
biology of these conditions or
established group within the University
from my previous work in Boston, which
screen compound libraries for
of Parma’s Department of Medicine
identified a modulator of a transcription
and Surgery aims to address this issue,
factor which is frequently mutated in
using a wide range of sample types to
TLL. We are now collaborating with a
investigate various aspects of leukemia
biotech company to validate a small
a ‘chemogenomics’ approach
and bone marrow transplantation.
molecule drug that inhibits this target,
to overcome this challenge,
Physician Scientist Dr Giovanni Roti
with the goal of translating these
working on a nanoliter scale
explained: “After spending several years
findings for clinical use.”
to identify new treatments for
at the Dana-Farber Cancer Institute
of primary patient samples. Without these precious
drug candidates. A group at the University of Parma is using
aggressive pediatric leukemias.
in Boston, USA, I returned to Italy last
On starting out, one of the lab’s first
year to set up a new unit for cancer
technology updates was to invest
research within the Hematology Unit
in a Tecan D300e Digital Dispenser,
here at the university. My work in Boston
which it now uses in several different
focused on pediatric cancers using
applications. “When we screen small
chemogenomics – systematic screening
molecules, we need to perform titrations
of small molecule libraries against
to find their IC50 values in multiple cell
individual drug targets or target families
lines or patient samples. Although
– to identify new targets and drugs for
this is possible to do manually in 96-
aggressive diseases such as leukemia
well plates, it is very time consuming
and neuroblastoma, and our work here is
to pipette 12 or 14 different dilutions
a natural progression of that.”
of up to 10 drugs for each cell type. The D300e simplifies this process
26
TECAN JOURNAL 2/2018
The Roti Lab takes a flexible approach
considerably, as well as allowing us to
to this research, adapting the techniques
work with smaller volumes in 384-well
it uses according to the goal of the
plates without the risk of pipetting
study and the samples available.
errors. It is also really fast, taking
Giovanni continued: “Sometimes we
between one and four minutes to set
start with a tissue sample, sometimes
up an entire 384-well plate depending
we use cell lines or cancer models.
on the number of compounds you want
But the basic process is always the
to test. We can also perform synergy
same; start by investigating the cellular
studies using this technology, which
biology, then use this understanding to
enables us to investigate many dilutions
identify and validate therapeutic targets
of two or more drugs in combination.
using both biochemical and genetic
This simply wouldn’t be possible
approaches, including CRISPR/Cas9
manually, as the time required for
knockdown if required. We are currently
pipetting would compromise the cells.”
DRUG DISCOVERY
ssentially, E we’re translating our pipeline from bench to bedside in a much shorter timeframe.
Andrea Gherli (left) and Giovanni Roti are using the D300e to screen small molecule drugs for AML
“The speed of the system also offers the
“Another feature we really like about
exciting prospect of being able to work
the Tecan D300e is that it is very user
To find out more about Tecan’s
directly with patient samples to help
friendly, and only requires minimal
inform therapies for the clinic. Our goal
training to set up even complex synergy
drug discovery solutions, visit
is to create a system for chemotherapy
studies. As a small laboratory, this is
drugs similar to the antimicrobial
a fundamental consideration when
sensitivity testing commonly used in
investing in any new technology.
clinical environments to guide antibiotic
With only two postdoctoral fellows
prescriptions. With the D300e, we don’t
and two PhD candidates – plus a
need a lot of time to set up the early
rotation of students from master’s
phase of these experiments; we can go
or undergraduate degrees – I cannot
from receiving the patient samples to
afford to have equipment that is not
testing compounds in the incubator in
accessible any time and to everyone.
just a couple of hours. If we happen to
The software automatically calculates
identify a molecule that is already in
how much of each compound is
clinical trials or for another indication,
required – and can randomize plate
we can ask permission from the local
layouts to avoid edge effects – which
ethics committee to use the drug in
means that inexperienced users don’t
patients. In a couple of cases, we’ve
have to perform a lot of calculations
actually been able to see a repeat
to work out the dilutions. Students
of what we’ve observed in vitro in
with limited laboratory experience
these patients, so it’s proving to be a
can therefore use this system without
good prediction of in vivo responses.
compromising the machine or risking
Essentially, we’re translating our
errors, which is a huge advantage,”
pipeline from bench to bedside in a
Giovanni concluded.
www.tecan.com/drugdiscovery To learn more about the University of Parma’s Department of Medicine and Surgery, go to
mc.unipr.it
much shorter timeframe.”
TECAN JOURNAL 2/2018
27
CLINICAL DIAGNOSTICS
Long-term partners in success Continuous advances in medical understanding are leading to rapid growth in the demand for diagnostic testing, making automation essential for laboratories trying to address this increase in their workloads. Many providers are also centralizing and consolidating testing to reduce costs, relying on instruments with higher throughput capacities to meet the demands of large service laboratories. Spanish biotechnology company Vircell recently partnered with Tecan to create a high throughput automated system for its VirClia® assays, building on the success of the company’s existing VirClia instrument. Vircell, based in Granada, Spain,
we have partnered with Tecan for
is very popular, providing on-demand,
specializes in the development and
the development of automated
automated testing for a wide range of
production of ready-to-use reagents
workstations ever since. Our latest
infectious diseases not covered by the
for clinical diagnostics. The company
system – the VirClia Plus, based
major diagnostics providers. This system
was founded in 1991 by two clinical
on a Freedom EVO® platform – is
is capable of processing 96 samples in
microbiologists looking to accelerate
a continuation of this relationship,
10 hours – which is sufficient to meet
hospital-based diagnostics for better
offering infectious serology testing
the clinical needs of most facilities – but
patient outcomes, and has grown into
in a high throughput format.”
very large laboratories can require two
a successful global player, with its
or three of these instruments to meet
products in use in over 80 countries across five continents. Vircell’s assay portfolio employs a wide range of technologies – ranging from traditional cell culture to molecular diagnostics – and focuses on esoteric infectious
Tecan has an excellent reputation in the IVD market, and you can always be sure of the quality of its solutions.
diseases. Joan Anglés, Marketing Director at Vircell Spain, described the
Chemiluminescent immunoassay
their throughput demands, and so we
company’s approach: “When Vircell
(CLIA) techniques provide an ultra-
looked to develop a higher throughput
was created, the aim was to produce
sensitive alternative to classical enzyme
instrument for the same monotest strips.
assays that simplified clinical laboratory
immunoassay formats, making them
Although we considered a number of
workflows. Quality and sensitivity
ideally suited to the detection of
laboratory automation suppliers to help
were key considerations, which is why
low abundance molecules, such as
us to develop this new platform, we
the company’s founders chose to
circulating biomarkers of infectious
have been working with Tecan for over
produce all the antigens needed for the
diseases. Vircell’s VirClia assays rely
20 years, and so the company was our
development of our kits in house. This is
on an indirect CLIA method for signal
logical choice of partner. Tecan has an
still true today, and ensures that we can
generation, with all the calibration and
excellent reputation in the IVD market,
guarantee the quality of our products, as
standards included in each test strip
and you can always be sure of the
well as be more flexible to the demands
to create a ‘monotest’ format. There
quality of its solutions. We first began
of the market.”
is no need to run separate calibrators
discussing the project at the AACC
or quality controls, and labs can avoid
exhibition in 2015, then worked closely
“Even in 1991, the demand for infectious
batch testing, allowing faster turnaround
with the Tecan team through to the
disease testing was growing rapidly,
of results using the company’s benchtop
instrument launch last spring.”
and so automation was considered
VirClia instrument.
essential to help laboratories manage
The result of this collaboration is a CE-
their workflow demands. Our first
The VirClia Plus system builds on
marked, fully automated system with
automated ELISA processor was
the success of the VirClia, as Joan
an on-deck capacity of up to 216 tests.
based on the Tecan Minilyser, and
explained: “Our original VirClia system
Using an eight-channel pipetting arm,
28
TECAN JOURNAL 2/2018
CLINICAL DIAGNOSTICS
the VirClia Plus can process 96 tests
has been excellent and, importantly,
in just three hours, with an integrated
this didn’t stop once the product was
To find out more about Tecan’s
Infinite® 200 PRO multimode reader
launched. Tecan’s engineers worked
Partnering Business, visit
to allow fully automated measurement
with our staff to ensure that the first
and reporting of results. It also offers
few instruments installed at customer
automatic barcode scanning, custom
sites performed exactly as expected,
assay set-up options and the ability to
which was vital to long-term success.
pipette directly from primary sample
Now that our engineers have become
tubes, providing a flexible, reliable and
more familiar with the system, we no
effective solution for diagnostic labs
longer require this kind of day-to-day
and hospitals.
support, but the company’s service
partnering.tecan.com To learn more about Vircell and the VirClia Plus system, go to www.vircell.com
team is always there if we come across “As with any complex project – and
something unexpected, which is very
especially any instrument intended for
helpful. We are also working with Tecan
the IVD market – there were a number
to add new features and capabilities to
of technical challenges that needed
the system, so it’s certainly an ongoing
to be overcome,” Joan continued.
relationship.”
“The support we have had from Tecan
The Freedom EVO-based VirClia Plus system offers walkaway processing of 96 samples in three hours
TECAN JOURNAL 2/2018
29
ENVIRONMENTAL TESTING
Detection at the molecular level Many common food additives and pharmaceuticals make their way directly into aquatic ecosystems. While their effects on humans are well documented, the impact on the environment and marine lifeforms is largely unknown. This has become the focus of a collaborative project involving researchers at the Karlsruhe Institute of Technology, which has adopted fluorescence polarization to explore the problem at the molecular level, screening compounds for their affinity to nuclear receptors. The Karlsruhe Institute of Technology
interactions on a nanoscale and their
of Tübingen, KIT and biotechnology
(KIT) in Germany was established in
applications for macro-level systems.
company Biametrics. The collaboration is
October 2009 following the merger
Siegfried Hohmann, a PhD student in
investigating the effects of food additives
of the Karlsruhe Research Center and
the analytical biochemistry group at
and pharmaceuticals – including their
the University of Karlsruhe, with the
IFG, said: “My research forms part of
metabolites and decomposition products
purpose of developing sustainable
Effect Net, a multidisciplinary project
– on aquatic ecosystems, as well as
solutions to tackle today’s societal,
on behalf of the Baden Württemberg
raising social awareness of the problems
industrial and environmental challenges.
Water Network, which brings together
these micropollutants can cause, in
KIT’s Institute for Functional Interfaces
around 20 researchers from across the
collaboration with consultancy firm
(IFG) studies solid matter surface
University of Heidelberg, the University
ÖKOnsult.”
Left to right: Sarah Roth, Siegfried Hohmann and Florian Durm with the Spark reader
30
TECAN JOURNAL 2/2018
ENVIRONMENTAL TESTING
Siegfried continued: “The aim of the project is to analyze the impact of these chemicals, from the molecular receptor level – which I am researching – through to the microbiome and behaviors of fish and snails. My particular focus is on four main compounds: the artificial sweetener sucralose, the anti-diabetic medicine metformin and
The platform can also read 384-well plates… which allows us to include our reference substance and three other unknowns in a single run, using 16 different concentrations in triplicate.
two antidepressant drugs, fluoxetine and citalopram. These substances are poorly digested by humans, so large
“We decided to use FP for fast screening
uses testosterone, which has a low
amounts end up in wastewater plants.
of samples, to identify compounds for
affinity for the estrogen receptor, and
Unfortunately, they are not effectively
more in-depth study by surface plasmon
zearalenone, which is an estrogenic
removed at this stage – further cleaning
resonance spectroscopy (SPR). We
metabolite that shows a strong affinity.
and filtration would be required – and
needed a platform that could offer
Results for varying testosterone and
enter the environment as a result.”
quick reads and, after looking at other
zearalenone concentrations were used
instruments on the market, we decided
to create a competitive plot, showing
“The nuclear receptor affinity of these
to purchase a Spark multimode
the relationship between concentration
different substances, their metabolites
microplate reader with enhanced Fusion
and the polarization of the emitted
and any products arising from UV
Optics. The system’s software is easy to
light. The half maximal polarization shift
decomposition can be determined
use, and my experience trying to set up
concentration (IC50) is characteristic
against reference standards with
the experiments on an older instrument
of each substance and its affinity, and
known affinity. A key technique for
meant I was already familiar with the
can be referenced against an estradiol
determining this affinity is fluorescence
way microplate readers work and which
standard. As expected, testosterone
polarization (FP). The principle of the
parameters could be optimized. The
showed no change in polarization as the
technique is fairly straightforward. A
platform can also read 384-well plates
concentration varied, while there was
sample is mixed with a receptor bound
– a deciding factor – which allows us
a clear decrease in polarization as the
to a fluorophore-labeled analog of the
to include our reference substance and
concentration of zearalenone increased.
compound of interest. If a compound
three other unknowns in a single run,
Overall, the technique has worked
present in the sample has a higher
using 16 different concentrations in
really well, and I’m looking forward to
affinity than the labeled analog, it will
triplicate. The readings can be taken in
adapting the method to support my
displace it and preferentially bind to
10 to 15 minutes – just enough to grab
research into micropollutants,”
the receptor. Polarized light is then
a bite to eat – in contrast to SPR, which
Siegfried concluded.
used to excite the fluorophore, and the
can take almost a day to give a single
difference in polarization between the
measurement, and is better suited to
exciting and emitted light is measured.
in-depth studies to determine kinetics
To find out more about Tecan’s
The key variable is the speed of rotation
and time-resolved measurements for
Spark reader, visit
of the labeled analog; when bound to
compounds identified through
www.tecan.com/spark
the receptor, rotation is slow, but after
FP screening.”
®
displacement, rotation is much faster.
To learn more about the
The difference in polarization between
“We purchased the Spark at the
excitation and emission is therefore an
beginning of the year, and initially
indicator of the level of displacement –
validated an assay involving estrogen
as more labeled analog is displaced, the
receptors, which are inexpensive
emitted light becomes less polarized.”
and easy to stabilize. The assay
Institute for Functional Interfaces, go to www.ifg.kit.edu
TECAN JOURNAL 2/2018
31
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Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Günter Bolzern/Zürich www.bolzern.tv Editor in Chief: Tecan Trading AG, Antonietta Allocca Editor: kdm/UK www.kdm-communications.com Editor: UP THERE, EVERYWHERE/Sweden upthereeverywhere.com Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, hello@tecan.com, www.tecan.com Images that are not owned by Tecan have been reproduced with permission. To register for the Tecan Journal please go to www.tecan.com/journal © 2018 Tecan Trading AG, Switzerland, all rights reserved.
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Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law. In general, the trademarks and designs referenced herein are trademarks, or registered trademarks, of Tecan Group Ltd., Mannedorf, Switzerland. A complete list may be found at www.tecan.com/trademarks. Product names and company names that are not contained in the list but are noted herein may be the trademarks of their respective owners. For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative. This journal may contain reference to applications and products which are not available in all markets. Please check with your local sales representative: www.tecan.com/contact