Tecan Journal
Edition 1/2019
Life Sciences, Diagnostics and Partnering
NuGEN joins the Tecan family
A revolution in milk sample analysis
Exceptional analytical possibilities from a new cell culture environment
Correct identification of autoimmune diseases
Pages 8-9
Pages 12-13
Pages 18-19
Pages 22-23
CEO WELCOME
Welcome Dear Reader, We were delighted to further expand our complete solutions portfolio in August, with the addition of next generation sequencing reagents. Through the acquisition of US-based NuGEN Technologies (now renamed Tecan Genomics), we are now able to better serve this market segment, combining innovative NGS kits and genomic sample preparation solutions with our industry-leading automated workstations to provide dedicated workflow solutions. This expansion complements our existing range of complete solutions, which has grown significantly in the last few years, to include smart consumables, solid phase extraction devices and immunoassays for specialty diagnostics. This latest addition to the Tecan family further extends our capabilities and expertise in this rapidly growing market, and the combination of NGS reagents and Tecan platforms will help to accelerate a range of R&D activities. We will soon be launching our first automated workflow combining the NuQuantÂŽ chemistry with the FluentÂŽ Automation Workstation (pages 8-9), so watch out for details of this on The Helicase, our dedicated genomics channel. I hope you enjoy the issue, Dr David Martyr CEO
2
TECAN JOURNAL 1/2019
CONTENTS
Contents 2
Welcome
4 - 5
Is what you don’t know costing you? Why instrument utilization data could be your lab’s biggest asset
6 - 7
How can custom automation accelerate commercialization of biotechnology breakthroughs?
14 - 15 imely automation for ultra-sensitive T immunodiagnostics
8 - 9
NuGEN joins the Tecan family
10 - 11 Bacterial behavior offers insights into drug development
12 - 13
A revolution in milk sample analysis
14 - 15
Timely automation for ultra-sensitive immunodiagnostics
16 - 17 Post-mortem RNA sequencing revolutionizes neurological research
18 - 19
Exceptional analytical possibilities from a new cell culture environment
26 - 27 Automating genetic screening to support expansion
20 - 21 Finding a clinical future for HMGB1? 22 - 23
Correct identification of autoimmune diseases
24 - 25 Combining smart technology with clinical expertise
26 - 27 Automating genetic screening to support expansion
28 - 29 Automating protein purification 30 - 31
Designer chemotherapy
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 1/2019
3
LABORATORY AUTOMATION
Is what you don’t know costing you? Why instrument utilization data could be your lab’s biggest asset As we move into the 2019 budget cycle with signs of a global economic slowdown on the horizon, laboratory administrators are no doubt feeling the heat. A combination of poor forecasting, inefficient use of resources, and a sudden economic downturn could create the perfect storm to capsize operations. Despite these high stakes, critical decisions about budget allocation, expensive equipment purchases, workflow optimization and cost-cutting strategies are often made based on incomplete information or even pure guesswork about laboratory asset utilization.
How much do you really know about
USD or more. Larger organizations, such
it is virtually impossible for a lab to get
how and when your instruments and
as clinical reference laboratories, contract
the most out of equipment investments.
consumables are being used? Could
research organizations and pharmaceutical
Your lab could be wasting an inordinate
better use of existing data be the key
companies, can be managing extensive
amount of time and money without
to overcoming this recipe for disaster?
fleets of high-end robotic systems
anyone even realizing it. For example,
deployed globally, bringing the total
you might:
In our uncertain economic climate,
closer to one billion USD.
laboratory managers and administrators
• Purchase instruments and equipment that are not really needed
are constantly challenged to minimize
Given these large investments, and the
risk and squeeze more out of their
critical nature of the work they support,
hard-won budgets and resources,
it is sometimes surprising how little is
unplanned maintenance or by having
even at the best of times. ‘More, faster,
understood about how equipment is
the wrong type of service contract in
better’ is the universal imperative that
being used in the laboratory. How many
place
finds its way into virtually everyone’s
hours per day is each system actually
performance targets. Year-on-year it
running, and what is the burn rate for
gets increasingly difficult to further
associated consumables, such as pipette
streamline operations and come up with
tips and sterile tubes? How could
new strategies to drive cost efficiency.
scheduling and workflows be adjusted
Having an inaccurate picture of how
to increase lab productivity? Which
turnaround times arising from
laboratory equipment and consumables
robots are performing sub-optimally?
inefficient use or deployment of
are used can result in missed
When should they be replaced, retired
equipment and resources
opportunities and costly mistakes.
or redeployed?
Laboratory asset management: the high
Accurate answers to questions like these
cost of not knowing enough
are vital for optimizing productivity,
A typical R&D lab may have two or
coming up with effective cost-cutting
three pieces of capital equipment per
strategies, and making sound capital
scientist, totaling upwards of one million
investments. Without this information,
The Blog 4
• Incur excessive service costs from
• Experience a disruption of operations when equipment fails or requires an unplanned service visit • Encounter bottlenecks and long
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 1/2019
LABORATORY AUTOMATION
Instrument utilization data could be
Improve budget forecasting and
However, manually-recorded usage
your lab’s biggest asset
justification of capital requests – Usage
data may need to be collected from
So, although it doesn’t typically appear
data can provide important insights into
numerous logbooks around the lab,
on the balance sheet, instrument
which instruments are worth replacing,
and there’s no guarantee that these
utilization data could be your lab’s most
where new equipment could provide
records will be complete and accurate.
valuable asset. An analytics solution that
added value, and whether current assets
Instrument log files are typically
enables you to visualize and analyze
are providing the expected return on
developed for service engineers only,
instrument and consumables usage in
investment.
with data in a form that is difficult and time consuming to extract, import and
real time could fill in the gaps needed to make more informed decisions. For
De-risk operations and optimize service
example, it could provide the missing
intervals – Usage and performance data
piece of the puzzle needed to:
is an important element of failure modes
The situation can be even more unwieldy
assessment to determine how often
when managing a fleet of systems
Increase efficiency and productivity –
critical equipment requires preventive
across multiple sites globally. In these
Historical instrument usage patterns
maintenance, and what level of service
cases, it is not unusual for the various
may highlight capacity gaps, help
coverage is most cost efficient.
labs to have different ways of working,
parse into a centralized database.
incompatible information management
you minimize downtime, and improve troubleshooting. For example, you
Why isn’t evidence-based laboratory
systems, and varying degrees of
might discover that instruments are
asset management the norm?
diligence. Even in the best-case
underutilized at the beginning of every
Given that the best decisions are
scenarios, the data may not be available
shift, or on a particular day of the week.
data-driven, why is laboratory asset
in real time, which can prevent rapid and
Periods of overuse might be associated
management still typically more of an
responsive decision-making.
with higher failure rates, or indicate the
art than a data-driven science? In the
need for an additional instrument or
vast majority of cases, the problem
Making it better
redeployment of existing equipment to
arises not from lack of data, but rather
Despite these challenges, advances in
ease the workload.
from the difficulty of aggregating and
software and cloud technology promise
analyzing the data. Most labs actually
to ease the burden of usage data
Enable more rapid responses –
capture a significant amount of useful
aggregation and analysis, enabling lab
Real-time usage data can give
data in handwritten logbooks, laboratory
managers to make better use of the
administrators instant visibility of
notebooks, instrument log files, and
wealth of historical data available, and
throughput and capacity for scale-up
service records. User ID, event details,
to gain real-time insights into exactly
across an entire fleet of instruments,
run times, error logs, and maintenance
how and when their instruments and
enabling more timely and effective
test results are all useful sources of
consumables are being used.
decisions.
information for trend analysis and troubleshooting.
www.tecan.com/blog TECAN JOURNAL 1/2019
5
AUTOMATION
How can custom automation accelerate commercialization of biotechnology breakthroughs? You’ve done your testing on the benchtop and proven that your new biotechnology innovation works in your hands. Now comes the exciting part – turning your solution into a breakthrough product that is ready for broader use and commercial launch. To get there, you need to optimize your processes so that you can ensure they are robust, operate within defined tolerances, and facilitate scale-up. What’s the fastest and most efficient way to get this done so that you can focus on your next bioscience advancements?
Biotech innovation requires automation
Biotech innovation drives custom
innovation
engineering
Optimization requires changing
Your latest biotechnology innovation may
multiple parameters and generating
not conform to the existing laboratory
statistically-sound results based on
automation standards for consumables,
the use of many replicates. Scale-up
components, automatable modules or
for production demands speed,
software. Innovation does not develop
reproducibility, and reliability. Once you
within such constraints. Attempting
have protocols that work in your hands,
to build an automation solution using
it simply isn’t practical to continue with
components that don’t meet all of your
manual processes to optimize product
requirements can limit your flexibility
performance, validation procedures
as you try to optimize your workflow.
and production workflows, while still
Creating a de novo solution could
meeting your time-to-market targets.
divert your efforts and sacrifice your
Taking innovative concepts through
time-to-market. An automation solution
to commercial application is a job for
that combines both custom-engineered
automation. However, off-the-shelf
developments and existing products may
automation solutions may not be up to
enable you to optimize your workflow,
the task – especially when innovations
while maintaining the flexibility and
entail entirely new protocols.
speed you need during your final stages of product development.
The Blog 6
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 1/2019
AUTOMATION
Biotech automation solutions: moving beyond the ordinary When you have automation needs that
• Even a custom-configured instrument isn’t exactly what you need • You need innovation in automation
don’t yet exist as standard lab solutions,
to match the innovation in your
it’s time to consider collaborating with
biotechnology
dedicated laboratory automation system engineers who will work with you to
A custom-automation engineering
investigate, invent and integrate new
partner should listen to your needs and
approaches that will take you beyond
be able to offer the dedicated resources
standard options.
needed to work with you throughout the most dynamic phases of your project,
You’ll know it’s time to consider
to create fully optimized and tested
collaborating with a custom engineering
systems that are tailored to your unique
team when:
requirements. Collaborating with system
• You need to focus your expertise
engineers who will push the boundaries
on bioscience, not inventing lab
in laboratory automation can help you to
automation instrumentation
push the boundaries in biotechnology.
• You have unique application workflows
www.tecan.com/blog TECAN JOURNAL 1/2019
7
GENOMICS
NuGEN joins the Tecan family NuGEN Technologies – a specialist in
adoption of genomics in life sciences
NGS and genomic sample preparation
research, it expanded its product
products – joined the Tecan Group in
development into NGS library
September 2018, opening up exciting
preparation to address the unmet needs
new opportunities for integrated NGS
in genomics sample preparation. The
workflow solutions.
portfolio now offers over 20 NGS and microarray sample preparation
NuGEN Technologies is based in the
products, covering a wide range of
US and was founded in 2000 around a
sample types, including RNA and DNA
core sample preparation technology
extracted from fresh and FFPE
called SPIA (single primer isothermal
(formalin-fixed paraffin-embedded)
amplification), a method that provides
tissues, single cells, and liquid biopsies.
robust amplification of small quantities
8
TECAN JOURNAL 1/2019
of RNA, even for poor quality samples.
In September 2018, NuGEN joined the
In these early days, the company’s
Tecan Group to become Tecan Genomics,
focus was on solutions for microarrays.
bringing a wealth of NGS reagent
Then, with advances in NGS and wide
knowledge to existing expertise in
GENOMICS
automation solutions. Lin Pham,
in seconds. Joining Tecan will help us
Enrique Neumann, Product and
Vice President of Research and
to contribute even more to the market,
Application Manager Genomics at
Development, has many years’
pulling together the expertise from
Tecan, commented: “This acquisition
experience in the life sciences sector,
both companies, and combining our
offers real value to our customers
specifically in genomics, and has
respective reagents and platforms.”
and their research. The combination
overseen product development at the
of NGS reagents and Tecan platforms
company since the NGS revolution.
The first joint solution is already
will support the R&D process, and the
She explained: “It’s amazing to see
under development, integrating and
collective expertise of the Tecan team
how things have advanced over my
automating NuQuant so that customers
will contribute to the development
time here; we have collectively learned
can pool libraries and load them onto
of combined solutions for every lab.
a tremendous amount, and apply this
sequencers without going through
We have prioritized a number of
knowledge to the development of
tedious quantification processes.
Tecan Genomics products that will
innovative products every day. Our
This new solution is based on a
be integrated and optimized for use
extensive knowledge covers all areas
standard NGS deck configuration of
on the Fluent system, providing our
of sample preparation, from DNA- and
the Fluent® Automation Workstation,
customers with a one-stop shop for
RNA-seq to target enrichment and
with prevalidated protocols to integrate
chemistry and instruments, supported
methylation studies. We are very much
NuQuant seamlessly with the Infinite ®
by one partner. The combination of
a customer-centric organization, and
plate reader. Once the DNA or RNA
innovative NGS products with our own
spend a lot of time interacting with
samples and reagents are loaded, the
industry-leading automation platforms
customers – attending conferences,
NGS libraries are automatically
will only help to accelerate and
visiting their labs and developing close
generated and pooled, resulting in a
empower research – it’s a perfect
relationships with them. Everything we
finished library pool for sequencing.
match.”
do is driven by understanding and
oining Tecan will help us to contribute even J more to the market, pulling together the expertise from both companies, and combining our respective reagents and platforms. anticipating their needs, and regular
Lin added: “Customers are very
feedback helps us to identify common
excited about Tecan Genomics; the
problems and develop solutions for
combined solutions will improve
them. The biggest bottleneck we
workflow efficiency and simplify
currently see in NGS sample prep is
communication – they only need to
converting nucleic acids into NGS
approach one company for reagents,
libraries – a vital step because the
automation services and application
ability to sequence well depends on it.
support. At the same time, they will
This has been our most recent focal
still have the option to use the
point, resulting in NuQuant ®, a novel
reagents on other platforms, and other
method that accurately measures
kits on Tecan systems.”
To find out more about Tecan Genomics, visit www.tecan.com/ngs or www.nugen.com
molar concentrations of NGS libraries
TECAN JOURNAL 1/2019
9
MICROBIOLOGY
Bacterial behavior offers insights into drug development Studying the co-operative and
The Department of Plant and Microbial
microbes is that we can observe their
competitive behavior between
Biology at the University of Zurich,
evolution in real time, using molecular
bacteria offers potential insights
Switzerland, brings together plant
tools to study how they evolve, behave
biologists, molecular biologists and
and interact with each other.”
into treating polymicrobial infections. Researchers at the
microbiologists to explore a diverse range of fields, from lake ecology to
“Our primary focus is understanding the
University of Zurich are using a
plant-microbe interactions and
co-operation and competition between
combination of biochemical and
pathogenicity. The 180-strong
microbes. For example, bacteria
gene expression assays to probe
department is home to Professor Rolf
voluntarily secrete compounds to create a
these community interactions,
Kümmerli and his research group, which
shared pool of metabolites, which we call
focuses on microbial evolutionary
‘public goods’. The entire group benefits,
ecology. He explained: “My background
because co-operation accelerates the
is in evolutionary biology and, following
uptake of nutrients for everyone in the
a PhD studying social interactions
community. These contributions are not
among insects, I decided to apply the
always equal though, and some microbes
concepts I had learned to microbes. One
– the ‘cheats’ – do not add anything to the
of the key benefits of working with
public goods pool, taking advantage of
hoping to better understand how microbial species evolve, co-exist and cause disease.
For research use only. Not for use in clinical diagnostics. 10
TECAN JOURNAL 1/2019
The Department of Plant and Microbial Biology at the University of Zurich, led by Professor Rolf Kümmerli (back row, far right)
MICROBIOLOGY
the system. There is also the potential
we can use specific compounds to
We now have an entire strain library we
for competition or ‘in-fighting’, with
disable the function or expression of
can rapidly screen in this way to see
bacteria secreting various toxins to give
these siderophores, potentially
how bacteria react to treatment, which
themselves an advantage. Many of these
preventing bacterial growth.”
generates a lot of kinetic data for
shared metabolites and toxins are
analysis. This type of assay is on a
responsible for the symptoms we
“We have two plate readers – an Infinite®
experience during an infection, and are
M200 PRO and an Infinite M Nano+ –
known as virulence factors. From an
which are key to carrying out our
“The support from Tecan has been very
ecological perspective, it’s fascinating,
screening assays. I used a Tecan reader
good. If we have any questions, we just
we’re interested in how these
at my previous workplace – where we
call the Tecan team, and they either
co-operative and competitive
had multiple platforms from a number of
explain things over the phone or –
interactions add up to explain how
suppliers – and it was clear to me that
because we’re right on their doorstep
species co-exist and cause disease.”
the Infinite 200 was the most
– they send an application specialist to
appropriate for this research. We bought
help us out; we’re very lucky. This kind of
“We have a number of projects studying
our first system in 2013, and it has been
support is important in an academic
these interactions both within and
ideal for these high throughput assays. It
setting, as people come and go in the
outside hosts. These polymicrobial
was soon running 24/7, with some users
lab, meaning that there is a risk of losing
infections are important to investigate,
having to wait a long time to run their
some technical expertise with the
as it’s still unclear what happens when
experiments, so we purchased the
equipment over time.”
microbes interact with one another, and
second system at the beginning of this
the underlying reasons for an increase in
year to cope with the volume of assays
“In the future, we would like to develop
infection severity. Ultimately, we want to
we wanted to perform.”
assays for polymicrobial infections to
know if we can steer these microbial
totally different level!”
monitor multiple competing strains – each with a distinctive signal – allowing
e now have an entire strain library we can W rapidly screen in this way to see how bacteria react to treatment, which generates a lot of kinetic data for analysis.
us to track community composition in real time. One of the key challenges is that many of these bacteria secrete multiple autofluorescent metabolites into the media, which interferes with detection of the genetic markers, but this is a technical issue that we hope to
population dynamics to reduce damage
“Our simplest assays use the readers to
solve. Ultimately, our long-term goal is
to the host. This has led us on to
measure optical density, helping us to
to see our research adopted for clinical
screening compounds to see whether
monitor the growth of bacteria treated
applications,” Rolf concluded.
they inhibit communication systems
with compounds inhibiting bacterial
between bacteria, which control the
communication. Conveniently,
expression of public goods and toxins.
siderophores exhibit autofluorescence,
To find out more about Tecan’s
One particular trait worth exploring
so we can easily measure production of
cell biology solutions, visit
concerns iron-scavenging siderophores.
these virulence factors with the
Typically, bacteria cannot grow in our
systems’ monochromators. In addition,
www.tecan.com/cellbiology
bodies because iron is not readily
we have constructed modified strains,
available. However, some bacteria
tagging the genes of interest with
secrete siderophores that extract iron
green or red fluorescent proteins to
from host cells. In vitro experiments – as
allow us to quantitatively track the
well as some in host systems – show that
expression of virulence genes over time.
To learn more about Professor Kümmerli research, go to kuemmerli-lab.weebly.com
TECAN JOURNAL 1/2019
11
GENOMICS
A revolution in milk sample analysis Quality control of milk is
AGRANIS, based in Laval, specializes in
an indicator of quality, since a high SCC
important to ensure compliance
milk analysis on behalf of dairy farmers,
is typically the result of an increase in
with regulations and to support
livestock consulting companies and vets.
white blood cells in response to an
Founded in 2005, the company now
infection. Generally, the SCC is
employs around 30 people and is
determined by flow cytometry, requiring
growing its services, as Genomic
analysis of individual milk samples from
analyzing multiple milk samples
Manager Dr Florent Perrin explained:
each cow in a herd, which involves a
from many individual cows is a
“We work mainly with milk samples,
large number of samples. However, prior
time-consuming process. French
performing biochemical and
genotyping of cartilage samples from
laboratory AGRANIS is using a
microbiological analyses to determine,
each cow in a herd makes it possible to
for example, protein or diuretic content,
determine the SCC of each animal in a
as well as carrying out ELISA tests and
single bulk tank milk sample.”
dairy farmers in their herd management. However,
new, automated genotyping technique to analyze bulk tank
qPCR. We process around 4.5 million
milk samples, saving time and
samples per year from across the
money on its testing services.
country, and focus on two primary
Genocellules® (Groupe Seenergi) – is
activities. Firstly, we assess the quality of
rather groundbreaking in agriculture,
milk from farmers according to industry
and relies on reliable DNA extraction
regulations and, secondly, we support
from both milk and cartilage samples.
dairy farmers and livestock consulting
This was not an activity we had done
companies to improve herd
before, and so we decided to purchase
management by determining the
a Fluent® Automation Workstation in
somatic cell count (SCC) for each cow.
October 2017 to help us achieve
The SCC is quantified as the number of
consistent results and cope with the
cells per milliliter of milk, and is used as
number of samples we would be
Members of the AGRANIS team with Dr Florent Perrin (right)
12
TECAN JOURNAL 1/2019
“This patented approach – called
GENOMICS
processing. We already had a Freedom EVO® 150 liquid handling workstation and two Sunrise™ absorbance microplate readers, and so it made sense to add a Fluent so that we could consolidate service and support. We had also had good reports about its reliability and reproducibility from other laboratories, which made us
utomating DNA extraction has saved us A time, and the enhanced speed allows us to achieve reliable batch processing of 96 samples in just two hours, running several plates a day.
even more confident of our decision. The Tecan team installed the platform and provided training, allowing us to
shaking and homogenization of samples,
us to achieve reliable batch processing
create our own programs.”
as well as heating buffers up to 70 °C.
of 96 samples in just two hours, running
We also have a Te-VacS™ module to
several plates a day. Today, we can
“Our workflow is separated into two
perform filtration steps without needing
analyze around 1,300 samples a week,
parts – the first step is to extract DNA
an external centrifuge, and have
rapidly genotyping each cow of a herd,
using a Nucleospin® Tissue 96 Kit from
developed a custom suction force profile
then determining individual SCCs from a
Macherey-Nagel. The two matrices
to optimize the process. Automating the
bulk tank milk sample. This is a real
present different challenges; the
protocol would not have been possible
benefit to our clients, as they only need
cartilage samples are processed in a
without the Robotic Gripper Arm™,
to routinely supply us with bulk tank
non-standard tube format, and the milk
which moves the plates and allows the
milk samples, and we can carry out
contains high levels of protein and fat.
workflow to be performed without the
twice as many analyses in the same
To overcome these issues, we developed
need for manual intervention. This
amount of time. I’m happy to say that
two separate protocols to handle each
set-up has given us high DNA extraction
the two protocols are running perfectly
sample matrix appropriately, benefitting
yields, which are essential for
and, to date, we have not encountered
greatly from the Fluent’s precise
genotyping. The entire extraction
any problems. We are very pleased with
pipetting, which allowed us to validate
process – lysis of samples, binding,
the Fluent’s performance, and are
the protocols within two weeks of the
washing and elution – also needed to be
already considering how we can develop
system being installed. Following DNA
carried out in such a way that we could
and program new protocols to support
extraction, genotyping is carried out on
carry out other procedures in parallel,
future projects.”
a Freedom EVO 150 liquid handling
which was straightforward thanks to the
workstation using a BovineSNP50 v3
user-friendly TouchTools™ interface. It
DNA Analysis BeadChip protocol from
guides us through the worktable set-up
Illumina.”
step by step – where to place different elements, number of samples, etc. –
To find out more about Tecan’s nucleic acid purification solutions, visit www.tecan.com/NAP
helping to minimize the risk of errors.”
“There were a number of experimental
To learn more about AGRANIS,
requirements that made the Fluent our platform of choice. We use a BioShake
“Automating DNA extraction has saved
3000-T elm (Quantifoil Instruments) for
us time, and the enhanced speed allows
®
go to www.seenergi.fr/en
TECAN JOURNAL 1/2019
13
OEM SOLUTIONS
Timely automation for ultra-sensitive immunodiagnostics High sensitivity immunoassays allow earlier detection and diagnosis of patient conditions, leading to earlier interventions and better patient prognoses. Immunodiagnostics company Singulex’s unique immunoassay technology allows early detection of even low abundance biomarkers for a wide range of conditions, and the company has partnered with Tecan to develop an intuitive, fully automated platform – the Singulex Clarity® system – which will allow clinical labs to take advantage of this novel technology for routine diagnostics.
The Singulex development group
Singulex is a California-based
sensitivity than other technologies and
immunodiagnostics company at the
is independent of any specific disease or
forefront of Single Molecule Counting
biomarker; we have already developed
technology. The company’s proprietary
assays for cardiovascular, infectious
technology provides unprecedented
disease and oncology applications.”
sensitivity, enabling the precise
14
TECAN JOURNAL 1/2019
measurement of biomarkers at
Single Molecule Counting technology
previously undetectable levels, offering
uses a familiar ‘sandwich-type’
a clearer and more definitive picture of a
immunoassay format that combines
patient’s health, as Dr Jeff Bishop,
capture antibodies immobilized onto
Senior Vice President of Diagnostic
paramagnetic microparticles with
Operations at Singulex, explained:
fluorescently-labeled detection
“Single Molecule Counting is a very
antibodies. However, this is where the
sensitive optical system that allows us to
similarity ends, as Jeff highlighted:
analyze low abundance biomarkers in
“After a wash step to remove unbound
blood samples in an extremely precise
assay components, we disrupt the
way. It offers up to 1,000-fold greater
antibody-antigen complex to yield free,
OEM SOLUTIONS
dye-labeled detection antibodies in
‘sample in, result out’ workflow that
an optically-inert buffer solution. By
could be deployed at customer sites.”
removing the analyte and magnetic particles, this significantly reduces
“We had two main goals for this
potential sources of assay ‘noise’,
project,” Jeff added. “The first one was
allowing individual fluorescently-labeled
to get this system into the market as
molecules to be detected as they pass
quickly as possible, and the second was
through the narrow interrogation
to ensure the same level of assay
window of a confocal detection system.
performance as in our clinical lab.
This method doesn’t rely on the total
Tecan’s engineers had already created a
fluorescence within the sample, and so
number of complete clinical platforms
we’re able to detect low abundance
based on the Freedom EVO® system,
biomarkers – such as proteins and
and so we felt that, with this experience,
nucleic acids – down to the femtogram
they could help us get to market more
per milliliter level.”
quickly than anybody else. We already had more than 10 Freedom EVO
The unprecedented sensitivity of Single
platforms in the laboratory, so we were
Molecule Counting enables much earlier
familiar with Tecan and the technology,
detection of time-critical biomarkers –
and knew the assays could be
such as the cardiac biomarker troponin
automated on this system. Because
– or bacterial infections, for example
Tecan has a portfolio of detection
C. difficile, than would be possible with
instruments, we were also confident that
traditional immunoassays. This allows
the company had the expertise to
appropriate care to be initiated much
replicate the design of our reader and
sooner. “By the time other systems can
incorporate it into the system. This took
detect these biomarkers, the patient is
a lot of the risk out of the instrument
really sick,” Jeff continued. “Single
development program.”
Molecule Counting can effectively identify much lower concentrations of
“When we first started out, we thought
these diagnostic markers much sooner,
that as much as 80 % of the system
leading to earlier intervention and better
would use off-the-shelf Freedom EVO
patient outcomes.”
parts, but it became more customized over the course of the development. Our
e were familiar W with Tecan and the technology… this took a lot of the risk out of the instrument development program. interface to guide the user through each step of the workflow.” “During the development process, we had weekly meetings with the Tecan development team; we visited the company’s sites in both Switzerland and Austria several times, and Tecan engineers came to our lab here in California. There were very few days when somebody from Tecan wasn’t talking with somebody from Singulex. Tecan isn’t just our development and manufacturing partner though. We now have a service relationship with the company, whereby Tecan’s engineers will be helping us to service the systems that are placed out in the field, ensuring support is available for the global Clarity landscape,” Jeff concluded.
Singulex began using the early detection
technology uses a unique, eight-well
capabilities of this technology for clinical
reaction vessel, so the gripper fingers,
applications in 2010, through a service
pipetting heads and integrated
To find out more about
provided, at the time, by its CLIA-
HydroFlex™ washer had to be modified
certified laboratory. “Physicians all over
to accommodate our consumables. The
partnering with Tecan, visit
the US would send blood samples to us,
system also has a four-channel Air LiHa™
and we ran them in our lab and reported
pipetting arm, a Robotic Manipulator
the results. In 2012, we decided to
Arm™, a MIO™ incubator and a shaker.
develop a fully automated platform with
The software is based on Freedom
and the Clarity system, go to
ready-to-use reagents – the Singulex
EVOware , but uses a custom,
www.singulex.com
Clarity system – creating a streamlined
touchscreen-based graphical user
®
partnering.tecan.com To learn more about Singulex
TECAN JOURNAL 1/2019
15
NEUROLOGY
Post-mortem RNA sequencing revolutionizes neurological research CNS disorders are often seen as the greatest area of unmet medical need, and are characterized by changes or degeneration in specific subsets of neurons and other cell types in the brain. Pharmaceutical start-up Cerevance is using a new sequencing method to study specific cell types from post-mortem brain tissue, helping the company to understand the pathology of these conditions in more detail than previously possible.
Cerevance was established in 2016 as an
and Cell Engineering at Cerevance,
individual neuronal cell types from
independent spin-out of global
explained: “A huge proportion of people
post-mortem human brain samples, so
pharmaceutical company Takeda. The
end up suffering from CNS diseases, and
that we can understand their role in
company is headquartered in Boston,
there are limited therapies on the market
different disorders. This method enables
US, but R&D takes place at its UK facility
that address the majority of them.”
us to work with thousands of isolated
on the Cambridge Science Park, and is
nuclei at a time, providing a deep
focused on neurological conditions such
“We use a novel technology – Nuclear
sequence that is a much higher
as Alzheimer’s, Parkinson’s, Huntington’s
Enriched Transcript Sort Sequencing
resolution than single cell or single
and some psychiatric disorders. Steve
(NETSseq™), developed in the
nucleus sequencing, and allowing
Sheardown, Head of Molecular Biology
Rockefeller Institute – to sequence
detailed transcriptomic profiling of
The Molecular Biology and Cell Engineering team (left to right): Steve Sheardown, Daniel Barker, Samuel Russell, Clare Bender and Jason Lawrence
16
TECAN JOURNAL 1/2019
NEUROLOGY
I t gave us the most consistent results, from the lowest amount of material, even using very degraded samples.
defined cell populations. We can use this
Steve added: “We evaluated all of the kits
data to identify functionally significant
available from different manufacturers for
changes to neuronal or glial cells during
extracting RNA and producing
a particular disease process, and to
sequencing libraries, and trialed several
assess the potential of new therapeutic
different kits against each other before
targets. We receive samples from brain
deciding on the Trio RNA-Seq library
banks across the US, UK and Europe, and
preparation kit from Tecan. It gave us the
this network has allowed us to build an
most consistent results from the lowest
extensive collection of tissue samples,
amount of material, even using very
covering all stages of different diseases,
degraded samples. The built-in DNA
as well as unaffected controls, generating
removal step at the beginning is very
well-defined patient cohorts. We can
effective, which is important because
identify critical differences between the
nuclei have a very high DNA to RNA ratio.
neuronal circuits of affected and
The kit saves us from doing this step
unaffected groups to build hypotheses
separately, which streamlines our
around what could be modulated to
workflow. It was also automation friendly
affect the progression of a disease,
and could easily be adapted to our
before moving on to drug discovery.”
current robot, so it met all of our needs.”
“We wanted to automate our workflow
“Tecan sent one of its technical specialists
and convert it into a high throughput
to us for a week to help us set up the
platform because, although the NETSseq
automated workflow. This was useful,
method was effective when first
because it’s quite complex to go from
implemented, we were conducting
tubes on a bench to plates on a robot,
everything manually. Our throughput
especially as our workflow has multiple
needed scaling up to be able to process
steps from RNA preparation to individual
larger numbers of samples to grow our
sample dilution. Working closely with
database. The challenge is that the
them helped us to optimize all of the
quality of post-mortem tissue can vary
small details, like reagent supply or tube
substantially between samples, and you
fill volumes, and the platform is now
can be working with highly degraded
running beautifully. It effortlessly copes
material, so we needed a solution that
with our current throughput – determined
could cope with that.” The recent
by the amount of brain material that can
acquisition of NuGEN by Tecan has
be processed and supplied to us – and is
brought a wealth of expertise in NGS
scalable so that we could easily cope
reagents and innovative products to the
with more, giving plenty of room for
Cerevance does, go to
Tecan family, and it was this expertise that
growth. The new set-up saves us a lot of
www.cerevance.com
led Cerevance to choose a Tecan solution.
time, while improving the consistency
and reproducibility of our workflow. Automation makes sample errors a thing of the past, and eliminates the possibility of a human transposing a row or making an error, which would lead to interpreting the wrong sequences for the wrong samples. Every tube and plate is barcoded and tracked by the robot, so we never lose a sample and can easily pull up its data on a spreadsheet.” “We’re now trialing the higher sensitivity Ovation® SoLo™ RNA-Seq System from Tecan, which will be beneficial for analyzing samples that are extremely degraded, or for looking at rare cells which are only present in low numbers. We’re also discussing other methods that we are interested in incorporating into our research, such as automated single nucleus sequencing. The relationship we have with the Tecan team is excellent; they are very easy to work with and have completely understood our needs from the beginning. I’m excited for the future,” concluded Steve.
To find out more about Tecan’s RNA-Seq solutions, visit nugen.com/products/rna-seq To learn more about the work
TECAN JOURNAL 1/2019
17
CELL BIOLOGY
Exceptional analytical possibilities from a new cell culture environment Researchers at the Grenoble
Biomimetic coatings have gained
strategy. We have developed an osteo-
Institute of Technology are
popularity in recent years as a promising
inductive coating that actively promotes
tool for biomedical cell culture
the regeneration of bone, aiding the
applications, thanks to the wide range of
healing process, and we are now using
available ‘building blocks’ and their
this to understand the molecular
potential for the localized delivery of
mechanisms behind the differentiation
has developed an innovative,
bioactive molecules. Professor Catherine
of stem cells into bone and related
automated layer-by-layer
Picart and her group in the Materials and
tissues, such as fat and cartilage.”
production technique, allowing
Physical Engineering Laboratory
high throughput in vitro
(Laboratoire des Matériaux et du Génie
The use of layer-by-layer (LbL) assembly
Physique, LMGP) – part of the Grenoble
techniques is becoming increasingly
Institute of Technology, France – are
common for the production of these
investigating this technology for the
biomimetic surfaces for in vitro
regeneration of bone tissue, looking
investigations, but the generation of
specifically at difficult clinical situations
surfaces involving costly and rare
where bone does not regenerate as it
biomolecules remains problematic.
should. She explained: “Most of the time,
Catherine explained: “LbL films are
a patient has the ability to self-generate
nanometer thin – they cannot be seen
their bones, or surgeons can find
by eye – so they are extremely fragile.
solutions. However, some injuries or
They need to be produced in a
conditions require a more innovative
consistent and homogeneous manner to
exploring the potential of biomimetic coatings to aid tissue regeneration. The group
investigations into the ability of these biomimetic coatings to control stem cell differentiation.
The Bioactive Coatings team at LMGP (left to right): Paul Machillot, Catherine Picart, Laura Clauzier and Jie Liu 18
TECAN JOURNAL 1/2019
CELL BIOLOGY
promote consistent and controlled cell
the well, followed by a wash step, before
growth, which is very difficult to achieve
the next layer can be added. Calculating
manually, especially as some LbL films
the exact volume that needs to be
have up to 24 layers.”
aspirated after adsorption is critical to ensuring homogeneity of each layer, as
Current techniques require milliliters
excess material will form a meniscus
rather than microliters of soluble
around the edge of the well. The
biomolecules, and are poorly adapted to
microplate is tilted during aspiration to
microplate-based applications. In
minimize the dead volume in each well,
addition, manual LbL coating of
and the ability to precisely control the
microplates is both labor intensive and
position of the pipetting head is vital to
user dependent, making it poorly suited
avoid damaging the film. These complex
to high throughput screening strategies.
liquid handling operations are managed
To overcome these issues, the LMGP
by Freedom EVOware, and we have
team decided to automate the LbL
supplemented the software with a
assembly process, harnessing the power
macro specifically developed for this
of a Freedom EVO® 100 liquid handling
novel application – a task that was far
workstation.1 Catherine continued:
from trivial.”
“Tecan’s Freedom EVO platform is well
he tools we have T developed could be of great value to cell biologists across the spectrum. on bone regeneration, this methodology would be just as useful for investigation of vascular systems, fat regulation and neurological diseases, where the presence of bioactive molecules are equally important for physiological functions. The tools we have developed could be of great value to cell biologists across the spectrum, and making these available to other
known in the biotech and
The optimal tilting angle and aspiration
pharmaceutical industries, and has been
volumes were determined with a series
used for many studies involving high
of experiments across a large number
throughput screening of cellular
of microplate wells, using fluorescently-
behaviors. The challenge for us was to
labeled films to provide a quantitative
References
develop a liquid handling protocol that
assessment of the film homogeneity at
would allow reliable, reproducible and
the bottom of each well. This not only
economical production of LbL films –
demonstrated the exceptional
something that had never previously
reproducibility of the technique –
been achieved in a microplate format.”
achieving CVs of less than 7 % – it also
1) Machillot, P et al. Automated Buildup of Biomimetic Films in Cell Culture Microplates for HighThroughput Screening of Cellular Behaviors. Advanced Materials, 2018, 30(27), e1801097.
scientists would be a dream come true,” Catherine concluded.
highlighted their suitability for high The exceptional control of liquid handling
resolution imaging, with good optical
offered by the Freedom EVOware®
transparency and a thickness of less
software – combined with the
than 10 micrometers. “Cells grown on
To find out more about Tecan’s
workstation’s ability to tilt the
biomimetic films can form cellular
cell biology solutions, visit
microplates during dispensing and
assemblies or aggregates, but they are
www.tecan.com/cellbiology
aspiration – is at the heart of the coating
not embedded within the matrix. This
protocol. “Each layer is deposited by a
makes it very easy to automate
To learn more about LMGP’s
simple physical adsorption process, but
spectroscopic and microscopic analysis,
the film is extremely thin and delicate,
particularly using Tecan’s integrated
research, go to
so it is essential not to disturb the
multimode readers, and allows us to
previously adsorbed layers. Once each
perform quantitative analysis of a wide
layer has been deposited, excess
range of assay types very quickly.
material needs to be aspirated out of
Although our team is currently focused
www.lmgp.grenoble-inp.fr
Freedom EVO with Tilting Rack is for research use only.
TECAN JOURNAL 1/2019
19
IMMUNOASSAYS
Finding a clinical future for HMGB1? Extracellular HMGB1 has been implicated in a wide range of conditions, from trauma and sepsis to epilepsy and lupus. As a result, this almost ubiquitous biomolecule is garnering increasing interest as a potential target for new therapeutics. In a recent review paper, Prof Ulf Andersson et al. summarized the latest advances in our understanding of the release, receptor interactions and functional consequences of extracellular HMGB1.
Professor Ulf Andersson
High-mobility group box 1 (HMGB1) is
effects of HMGB1. He explained why the
one of the most common nuclear
molecule has been implicated in so
proteins in the body, with around one
many diseases: “If you search the
million copies per cell. However, it is the
literature, there are at least 14 cellular
extracellular release of HMGB1 that is
receptors that have been identified as
generating significant interest in the life
potential HMGB1 receptors, but only two
sciences and biomedical sectors.
– TLR4 (Toll-like receptor 4) and RAGE
Considered the prototype damage-
(receptor for advanced glycation
associated molecular pattern (DAMP)
endproducts) – are well documented.
molecule, extracellular release of HMGB1
This widespread implication of HMGB1 in
is a molecular ‘alarm’ signal that triggers
various cellular pathways may be due to
inflammation and tissue repair. Ulf
its role as a transporter protein, binding
Andersson, a senior professor at the
to other immune-activating molecules
Karolinska Institute, has spent over 20
and facilitating their uptake into cells via
years investigating the inflammatory
the lysosomal compartment.”
HMGB1 transports extracellular LPS to caspase-11 in the cytosol HMGB1-LPS complex RAGE
Dynamin-dependent endocytosis Caspase-11 Endosome
Lysosome
TLR4
HMGB1-mediated membrane damage
Cytokines Pyroptosis Leaking lysosome
HMGB1 transports LPS into endothelial and myeloid cells causing Gram-negative sepsis 20
TECAN JOURNAL 1/2019
F
IMMUNOASSAYS
“One of the key features that
preclinical inflammatory disease models,
complicates our understanding of
making it a promising therapeutic target
extracellular HMGB1 activity is its ability
for both infectious and sterile
to permeabilize the membrane of the
inflammation. The next step is to
lysosome. Under normal circumstances,
translate these findings to a clinical
molecules taken up via the lysosomal
setting, and several promising
compartment are digested to avoid
humanized anti-HMGB1 monoclonal
disrupting cellular activity. However, in
antibodies are already under
the acidic environment of the lysosome,
development. Ultimately, HMGB1
HMGB1 causes the lysosomal membrane
antagonists could hold the key to the
to ‘leak’, allowing intact biomolecules to
treatment of conditions such as sepsis
be released into the cytosol. Depending
or arthritis, but there are a large number
on the nature of the co-transported
of other conditions that could also be
molecule, this can have a range of
targeted. As a proof of concept,
intracellular effects, creating a highly
focusing on a disease with a distinct and
complex picture of HMGB1 activity.
easily measurable outcome – such as
For example, in systemic lupus
post-sepsis anemia or acetaminophen-
be shipped around the world, but it also
erythematosus, HMGB1 transports
induced liver injury – may provide a
precludes the comparison of data from
dsDNA from dead cells into the cytosol,
better starting point. This will allow
different sites and studies. The
inducing type I interferon release.
clear, quantifiable assessment of the
commercial availability of the IBL HMGB1
Another class of molecules
potential benefits of reducing
ELISA, which is now being used in
co-transported into cells via this route
extracellular HMGB1 levels in a less
research applications globally, is
is lipopolysaccharides/endotoxins (LPS).
time-critical and clinically complex
transforming the way these studies can
Found in the outer membrane of
setting.”
be performed, allowing accurate
Gram-negative bacteria, HMGB1 binds
assessment of pre- and post-treatment
and transports LPS into endothelial and
“Regardless of the indication, the ability
extracellular concentrations. This has
myeloid cells via the RAGE pathway.
to accurately measure HMGB1 will be
generated some very exciting results at
Once released into the cytosol, LPS can
vital. Previous studies have been
the preclinical level and, as HMGB1 is so
lead to the life-threatening systemic
hampered by the lack of a standardized
well evolutionarily conserved – it is over
inflammation that occurs in Gram-
and reproducible assay, instead relying
99 % identical in all mammals – we have
negative sepsis.”
on tests developed in house by a few
a real hope that these therapies will be
research laboratories with a special
successfully translated into a clinical
“HMGB1 antagonists have already
interest in this field. This not only delays
setting in the not too distant future,”
been highly successful in a number of
analysis of samples, which often need to
Ulf concluded.
he commercial availability of the IBL T HMGB1 ELISA, which is now being used in research applications globally, is transforming the way these studies can be performed, allowing accurate assessment of pre- and post-treatment extracellular concentrations.
The full review article can be found at: Andersson, U; Yang, H; Harris H. Extracellular HMGB1 as a therapeutic target in inflammatory diseases. Expert Opin Ther Targets, 2018, 22(3), 263-277.
To find out more about IBL’s HMGB1 ELISA kits, visit www.tecan.com/immunoassays
For research use only. Not for use in clinical diagnostics. TECAN JOURNAL 1/2019
21
IMMUNOASSAYS
Correct identification of autoimmune diseases Rheumatoid arthritis, type 1 diabetes and celiac disease are familiar examples of autoimmune disorders that occur when the immune system attacks healthy cells or tissues by mistake. Myasthenia gravis is a far rarer autoimmune disease, causing weakness in the skeletal muscles. LAB Maastricht UMC+ in the Netherlands has a long history with myasthenia gravis, using gold standard immunoassays to enable accurate diagnosis and provide the information necessary to allow effective treatment.
LAB Maastricht is the amalgamation of
technology-based satellite labs – such as
validated the IBL International (Tecan)
eight diagnostic functions at the
flow cytometry, protein electrophoresis
ARAb RRA on the recommendation of
Maastricht University Medical Center
and advanced immunodiagnostics – for
a clinician colleague specializing in
(MUMC+), including the central
specialist testing.”
MG,” Jan added. “This assay uses
diagnostics lab, pathology, medical
receptors from human muscle in
microbiology and clinical genetics. Its
Jan continued: “We have a special
combination with a radio-labeled
1,000-strong team of laboratory
interest in myasthenia gravis (MG) here
alpha-bungarotoxin snake venom
specialists, clinicians, scientists and
at MUMC+ in terms of both clinical
marker. The venom binds very
technicians offers a broad range of
practice and research, including a
specifically to acetylcholine receptors
routine and specialist diagnostic services,
robotic thymectomy surgery program
without interfering with ARAb binding,
as well as performing validation of new
that was established after it was shown
so there’s no need to purify the
methods and supporting clinical trials.
that the thymus is involved in the
receptor. This is a very clever system,
production of autoimmune antibodies,
and the assay works very well, with
Dr Jan Damoiseaux is a laboratory
and its removal may be beneficial to
good reproducibility and sensitivity.
specialist in medical immunology at LAB
MG patients. We therefore have a large
It’s also quantitative, which is
Maastricht UMC+, and has specialized in
number of MG patients who come to us
particularly important for therapeutic
autoantibody diagnostics since 1999. He
for investigations, and it is vital that we
monitoring. Our neurologists aim for a
explained: “After my PhD and a couple
accurately diagnose their condition
50 % reduction in a patient’s
of years in immunology research, I was
through laboratory testing.”
ARAb levels after treatment with
offered a three-year training program
immunosuppressants. These therapies
here at MUMC+, covering all aspects of
Around 85 % of patients with clinical
take time to work, and so ongoing
laboratory investigations for
symptoms of MG are positive for
monitoring of ARAb allows the
autoimmunity, immunodeficiency,
autoantibodies to acetylcholine
treatment to be increased or changed
allergy, transplantation immunology and
receptors, a transmembrane protein
if it is not having the desired effect.”
malignancies of the immune system. I
present in muscle cells. Radioreceptor
am now responsible for autoimmune
assays (RRA) are the preferred lab
For the 15 % of patients with clinical
diagnostics in the lab – as well as
tests for the detection of acetylcholine
symptoms of MG but no detectable
teaching medical students and clinical
receptor autoantibodies (ARAbs) as,
ARAbs – known as seronegative MG –
trainees – and my research on
unlike ELISAs, these assays maintain
it is now known that around 40 % have
autoantibody detection systems is
the three-dimensional structure of the
antibodies against muscle-specific
very much linked to my clinical
receptor that is essential for antibody
kinase (MuSK), a protein involved in
responsibilities.”
recognition. “We originally trialed and
cross-linking the acetylcholine receptor
“Being a university hospital, we offer a broad spectrum of assays, with around 70-80 % of patient samples coming from our own hospital, and the rest from other regional hospitals and primary care. The lab has a central area for routine testing, supported by
22
TECAN JOURNAL 1/2019
ur previous experience with the O company’s assays gave us confidence in its performance, so we didn’t look at any others.
IMMUNOASSAYS
to the cytoskeleton. Jan continued: “The discovery of MuSK autoantibodies is very important, because these patients have a more severe form of MG. Therapy is therefore more intensive, most commonly with rituximab immunosuppression to deplete B cells. Unlike the 50 % reduction target for ARAbs, the clinical aim for these patients is to completely eliminate anti-MuSK antibodies, making a sensitive assay vital. Some other laboratories in the Netherlands only
Kathleen Mallet, Veerle Mengels, Caroline Bijnens, Jos Wiersma and Jan Damoiseaux from the specialist immunology laboratory at MUMC+
report qualitative results, but we believe that it’s important to use a quantitative assay, allowing clinicians to review the efficacy of therapy. Tecan makes the only commercially-available quantitative anti-MuSK ELISA, and our previous experience with the company’s assays gave us confidence in its performance, so we didn’t look at any others.” “We have a good relationship with Tecan, and I’ve presented my work at several user meetings. I’ve also set up discussions for some of my research colleagues, who are looking at acetylcholine receptor blocking antibodies, and Tecan is keen to help with their research,” Jan concluded.
To find out more about IBL’s MG assays, visit www.tecan.com/immunoassays To learn more about LAB Maastricht UMC+, go to www.labmaastricht.nl Agrin-induced clustering of acetylcholine receptors via MuSK TECAN JOURNAL 1/2019
23
CLINICAL DIAGNOSTICS
Combining smart technology with clinical expertise NGNY Devices is striving to fully automate laborious pre-analytical sample processing for clinical testing, helping to increase throughput and reduce human error. Its systems are already providing advanced sample sorting and aliquoting across Europe, offering a significant advantage over manual sample handling for busy clinical labs. Clinical laboratories routinely process
samples are processed quickly,
This is when our company truly evolved,
thousands of patient samples a day to
accurately and confidently. Joan
as the Cavro Omni Robot’s cutting-edge
aid in diagnostics, treatment and disease
Viladomat, Engineering Manager at
automation was perfect for our needs.”
prevention. Handling copious patient
NGNY, explained: “Our company started
specimens in a variety of tube formats is
building automation equipment in 2006,
“We quickly built a strong collaboration
time consuming and labor intensive, and
and our systems are now used in clinical
with the Cavro team, combining their
so many of these busy departments rely
laboratories across Europe. From the
liquid handling expertise with our
on streamlined workflows and
beginning, we have looked for ways to
understanding of clinical samples and
automation to enable efficient and
improve our existing products to better
workflows to fully automate the
error-free analysis. NGNY Devices, based
meet client needs. Many of our early
aliquoting process. Incorporation of the
in Spain, uses its engineering knowledge
customers repeatedly asked us if we
Cavro Omni Robot and Air Displacement
to develop automated sorting and
could develop instruments for
Pipettor (ADP) into our platforms
aliquoting systems, helping laboratories
performing liquid handling tasks and we
allowed us to create robust and flexible
to increase reliability and traceability,
were able to do this after discovering
systems. The Omni is a reliable OEM
and ensure that high numbers of patient
Tecan Cavro® components in 2010.
liquid handling robot and is incredibly
The AQUA8000 offers complete automation of pre-analytical sample handling, reducing errors and freeing staff to perform other tasks For research use only. Not for use in clinical diagnostics. 24
TECAN JOURNAL 1/2019
CLINICAL DIAGNOSTICS
easy to integrate; it was the perfect size and shape to fit within the dimensions of our instrument. The air displacement pipetting capabilities of the ADP offer precise liquid transfers from 0.1 to 1 ml, and this is much easier than our previous set-up, which required bottles of water as a system liquid. The combined ADP and Omni Robot allows single channel pipetting, including tip pick up,
e quickly built a strong collaboration W with the Cavro team, combining their liquid handling expertise with our understanding of clinical samples and workflows to fully automate the aliquoting process.
aspiration from a source tube, dispensing into a clean secondary tube, and ejection of the tip once finished. A built-in sensor is able to detect if a tip has not attached properly or fallen off, flagging any potential errors, which is a great feature for peace of mind.” This collaboration has led to the development of the fully automated AQUA product line of pre-analytical platforms, which sort, uncap, aliquot, recap and store tubes, reducing the risk of errors and freeing staff to perform other tasks. Joan continued: “When samples arrive in a lab’s specimen reception area, they can be loaded straight onto our AQUA systems, which register the tubes and sort them based on their destination within the laboratory. The instrument then aliquots samples as required, reducing the need for users to perform manual pipetting, and moves tubes from one rack to another, minimizing the chance of mistakes. The exact set-up is easily customizable by the user, allowing them
“Following the success of the AQUA7000 platform, we broadened our portfolio to include the AQUA8000, which allows more tubes to be processed at a time, increasing throughput and productivity. Tecan has supported us throughout the design and production of our systems, providing application support and answering any questions that arose. We were shown how to adjust the parameters of the robot, such as the length of the axes, which is incredibly easy to do, and the availability of 3D CAD files made the configuration process even simpler. Using Tecan Cavro components for the AQUA product line effectively gave us a
The AQUA systems can handle a wide range of tube formats
complete liquid handling solution, without having to build each individual module ourselves. We are very happy with both our relationship with Tecan, and the products that we have been
To find out more about Tecan’s
able to develop. We are continuously
Cavro components, visit
to change parameters – such as sample
expanding our product portfolio to
volume or label information – to suit the
anticipate our customers’ changing
needs of their laboratory workflow. It
needs, and currently have a new system
can also detect if there is an issue with
– incorporating the Omni Robot in a
pipetting, such as a clotted sample,
slightly different way – in the design
moving the sample to a different rack to
process, which is very exciting,” Joan
allow manual processing.”
concluded.
www.tecan.com/components To learn more about NGNY Devices, go to www.ngny.tech
TECAN JOURNAL 1/2019
25
GENETICS
Automating genetic screening to support expansion Genetic testing to screen for congenital defects is useful to identify susceptibility to, or the cause of, many diseases. The medical genetics department at the Policlinico di Milano – a teaching hospital in Italy’s Lombardy region – is using automation to improve the throughput of its genetic screening workflow, aiming to increase the number of diseases it can test for, and the number of samples it can handle.
Policlinico di Milano is one of Italy’s
or Parkinson’s. We also perform pre-
completely different set of genes in
oldest hospitals, founded more than 500
implantation genetic diagnosis (PGD)
each sample we receive, depending on
years ago. In addition to patient care,
for couples undergoing IVF, to ensure
the disease. For some, it’s one gene,
the hospital is heavily involved in
that embryos are free from any genetic
such as the CFTR gene for cystic
research in a variety of areas, from
diseases. However, the main bulk of our
fibrosis. For others, it’s a large panel,
neurology to cardiology. The medical
work is the screening of newborn
about 50 genes for Parkinson’s, for
genetics department is no exception,
samples, where access to early diagnosis
example. We also work with a lot of
carrying out studies to understand the
and treatment can be vital to long-term
different sample types including blood
role of mutations in disease, alongside
health. Samples are collected from all
and dried blood spots, buccal swabs,
its routine screening of patient samples
newborns born in the Lombardy region
chorionic villi, amniotic fluid, urine and
to diagnose diseases such as cystic
– about 90,000 births a year – and our
tissue biopsies. It’s therefore very
fibrosis, thalassemia and Duchenne
role is to confirm the biochemical
important to have a workflow that has a
muscular dystrophy. Manuela Seia, head
findings for patients who have already
high throughput and is incredibly
of the medical genetics laboratory at the
tested positive for disease biomarkers,
precise, so that we can get an accurate
hospital, explained: “We can identify the
uncovering the genetic cause.”
diagnosis with a quick turnaround time
causes of many conditions, from
from sample to result. We rely a lot on
microdeletions of the Y chromosome in
“We currently process around 5,000
NGS and have developed our own
male infertility to the multitude of genes
samples a year and our workflow is
molecular techniques to achieve a high
associated with Ehlers-Danlos syndrome
quite complex. We might be analyzing a
level of precision in a short time. We
The medical genetics team at Policlinico di Milano 26
TECAN JOURNAL 1/2019
GENETICS
use a NextSeq™ 550 platform (Illumina) for exome sequencing – to identify
he systems have been a great asset to us; T they have significantly reduced the amount of manual work we conduct – which was very laborious – and they save us huge amounts of time.
novel causative genes – and a MiSeq™ platform (Illumina) to perform a more
indexes for every sample. Introducing
panels. This will include amino acid and
targeted approach, looking at the
these index sequences onto DNA
vitamin B defects, fatty acid oxidation
association of a panel of genes with a
fragments enables 96 different samples
defects, galactosemia and
certain disease. We also use Sanger
to be pooled into a single lane on a flow
hyperammonemia, and
sequencing for thalassemias and
cell and sequenced together, and the
aminoacidopathies, such as
hemoglobinopathies, and to confirm
Freedom EVO allows this to be
phenylketonuria and maple syrup urine
the results of NGS analysis.”
performed automatically, reducing
disease. With these extra panels, we
error rates.”
predict that metabolic diseases will account for up to 30 % of our workload
Manuela continued: “Originally, we were performing all of our workflows
“The systems have been a great asset to
next year and, although this is a
manually, which took up a lot of time
us; they have significantly reduced the
significant increase in our throughput,
and was prone to error. Colleagues in
amount of manual work we conduct –
our staff numbers will remain the same.
another lab were successfully using a
which was very laborious – and they
I’m confident that our Freedom EVO
Freedom EVO workstation to
save us huge amounts of time. It’s also
set-up will easily support this expansion
automate their workflows and so, four
much more cost effective to automate
and that we will continue to achieve a
years ago, I decided to get two
the workflow, rather than hiring
high throughput, quick turnaround time
standard platforms, and then added
numerous people to perform everything
and precise diagnoses,” concluded
another system two years later. Each
manually. Not only that, but automation
Manuela.
Freedom EVO workstation is used for a
improves the quality of our results and
completely different application; the
reduces the chances of errors which
first is used for PCR set-up to test for
could potentially lead to misdiagnosis
short tandem repeats or mutation
of patients.”
®
To find out more about Tecan’s genomics solutions, visit
analysis. The second platform
www.tecan.com/genomics
automates the preparation of DNA for
“When the platforms first arrived, Tecan
Sanger sequencing, based on purifying
helped us to adapt them to our
PCR products; at this stage it is vital
workflow. The Tecan team was very
To learn more about the work
to remove all unincorporated dye
helpful and everything has worked
of Maneula at the Policlinico di
terminators from the PCR reaction to
perfectly since. The Freedom EVOs are
Milano, visit
prevent base calling errors. Our third
user friendly, faster than other systems
www.policlinico.mi.it/
Freedom EVO is used for NGS library
I’ve seen, and I think they offer the best performance on the market – we’re very
i-nostri-professionisti/
preparation – including Agilent SureSelect™ and HaloPlex workflows,
happy with them. Next year, we will be
and other assays based on MASTR
adding a panel of 85 genes involved in
technology – which can be complex,
pneumopathies to our screening, as well
particularly because we use specific
as further metabolic disease gene
profilo/601/seia-manuelabiologo-responsabile-di-unitoperativa-semplice
TECAN JOURNAL 1/2019
27
LIFE SCIENCES
Automating protein purification The initial screening of chromatographic conditions can be a major bottleneck in the development of protein purification protocols. Italian company BiCT – Biological and Chemical Technologies – has turned to automation to overcome this issue, implementing parallel processing of miniature chromatography columns on a liquid handling platform to enable rapid simultaneous screening of a range of conditions. This maximizes the prospect of achieving the best result for each project while saving time and resources. Lodi-based BiCT began life nearly a
Silvia continued: “One of our biggest
the speed of development of new
decade ago as a contract research
assets is automation. We offer a broad
protein purification protocols. “The core
organization (CRO) focused on the
range of automated high throughput
role of the automation lab in the new
pharmaceutical sector, establishing a
screening protocols for molecular
facility will be strain improvement and
core business developing biocatalysis
biology, fermentation, purification and
chromatographic purification, from
and biotransformation protocols
biocatalysis, developing efficient,
strain development to purification of
using enzymes and micro-organisms.
cost-effective processes. Automation
enzymes, important metabolites and
Subsequently, the company expanded
allows us to design our experiments to
proteins of pharmaceutical value.
its field of interest, applying the same
test numerous variables, increasing
Chromatography is a key step in our
technologies to other sectors, including
throughput and enabling many different
workflows, and we chose Tecan
nutraceuticals, cosmetics and
protocols to be evaluated to ensure we
because, in our opinion, it is the best
agrochemicals. More recently, it has
provide the best possible solution. The
system available.”
turned its attention to production, and
data is critically analyzed and forms the
now operates as both a CRO and a
basis of innovations that progress to
As a prelude to establishing a dedicated
manufacturing facility. Silvia Rapacioli,
the pilot plant and, ultimately, become
automation hub, BiCT worked with
Co-CEO and Marketing Manager of BiCT,
manufactured products.”
Tecan to explore the use of the Te-Chrom™ and Te-Chrom Shuttle with
explained: “We provide services to customers from all around the world
BiCT is set to move to larger premises in
Repligen OPUS® RoboColumns® on a
who appreciate our industrial focus. Our
the near future that will include a
Fluent® Automation Workstation for
services include everything from initial
dedicated automation unit. In
sample purification. “We initially
research all the way through
preparation for total laboratory
evaluated the RoboColumn technology
optimization to pilot-scale production
automation (TLA), the company is
on a Freedom EVO® workstation, and
and technology transfer to the
implementing, among other
have now implemented this application
customer’s own lab for use on an
applications, parallel processing of
on the Fluent. The system is very
industrial scale, with minimal
miniature chromatography columns on a
intuitive with straightforward software,
adjustments to the process.”
liquid handling workstation to increase
and allows us to automate parallel
[ Automation] gives us the opportunity to try as many options as the team can think of, exploring their creativity to its maximum potential, which increases success rates and the potential for devising innovative, ‘out of the box’ solutions.
For research use only. Not for use in clinical diagnostics. 28
TECAN JOURNAL 1/2019
LIFE SCIENCES
The BiCT team with the Fluent Automation Workstation
purification on miniature
“The success of our company depends
chromatography columns,” said Silvia.
on the satisfaction of our customers,”
To find out more about Tecan’s
Roberto continued. “With automation,
protein purification solutions, visit
Roberto Verga, Co-CEO and Business
we increase the chance of finding the
Development Manager, took up the
right candidate biologic and the best
story: “There are several things to
purification protocol quickly and cost
consider when you are developing a
effectively. Previously, we could only
purification protocol, such as the column
evaluate one column at a time. This was
volume, type of resin and screening
a huge effort, consuming a great deal of
conditions, which is a bottleneck when
time and resources, with the risk of
you can only run one column at a time.
missing the best results that can come
The big advantage of implementing the
from testing several conditions
RoboColumns on the Fluent is that we
simultaneously. A purification protocol
can run eight columns in parallel – 96 in
that used to take a month to set up can
one run – simultaneously studying a lot
now be established in just one week,
of parameters, including the kind of
which is a huge saving in terms of both
resin, flow rate, binding capacity, sample
time and resources.”
www.tecan.com/bioprocessing To learn more about BiCT, go to www.bict.it
clean-up and the best ratio of sample to resin. This allows us to speed up the
“The potential to integrate the entire
all-important screening process and
strain improvement workflow into our
move on to the next stage – process
lab automation is very important. It is a
scale-up – more quickly. The system is
real benefit not only for our customers,
very reliable, and we can easily change
but also for our staff. It is like having an
our protocols to use the larger volume
extra five or six collaborators that never
RoboColumns, which have proved
get tired. It gives us the opportunity to
representative of the subsequent
try as many options as the team can
industrial scale-up. We have also been
think of, exploring their creativity to its
able to engage with chromatographic
maximum potential, which increases
media manufacturers by, for example,
success rates and the potential for
comparing different resins, developing
devising innovative, ‘outside of the box’
specific resins suitable for customers’
solutions for every project. TLA will
applications, or evaluating the
undoubtedly be a key resource for
performance of new resins to generate
contract research in the future,” Silvia
data for technical brochures prior to
concluded.
the product launch.”
TECAN JOURNAL 1/2019
29
DRUG DISCOVERY
Designer chemotherapy
Brain tumors are the most common cause of mortality in childhood cancer patients, and treatment options are limited, especially for recurrent cases. Researchers at the University Hospital Düsseldorf are using automation to develop a more personalized approach to cancer medicine, performing rapid drug screening to identify novel therapeutic strategies to improve the chances of survival. Understanding of the underlying biology
clinically-established compounds, which
and 80 copies of our libraries per run –
and causes of tumor development is
provides a more comprehensive drug
helping to minimize the potential for
critical to advancing treatments and
response profile. We then use
batch effects and allowing us to
improving mortality rates. Researchers
bioinformatics tools to determine
investigate several tumor models at the
at the University Hospital Düsseldorf are
distinct drug classes or molecular
same time. It’s also very cost effective,
taking a holistic approach to this difficult
targets of interest. We try to manipulate
as we use a fraction of the materials that
area of oncology, combining genetic and
the biopsy-derived cultures as little as
we would consume if we were
epigenetic research with proteomics and
possible, to make sure they are similar to
conducting everything manually.
drug screening. Daniel Picard, Senior
the original tumor. Once we have
Originally, we were using 384-well plates
Scientist in the Pediatric Neuro-
harvested enough cells, we plate out and
but, more recently, we have begun using
Oncogenomics department, explained:
screen the tumor cells, using an in-house
1,536-well plates instead, which has
“Our main goal is to characterize
algorithm to determine IC50 values. This
turned out to be incredibly useful. We
tumor-specific alterations affecting the
data allows us to identify which drugs
performed several comparison studies
coding and non-coding genome to
are most effective against that individual
to make sure swapping formats didn’t
refine existing or develop novel
tumor model. After screening, we also
affect our results, and the correlation
treatment strategies. Our research focus
perform high resolution DNA
scores were very high, proving just how
is the epigenetics of pediatric brain
methylation analysis on both the tumor
reliable and consistent the platform is –
tumors – DNA methylation, microRNAs,
biopsy and the cultured cells, to identify
it’s amazing.”
and long non-coding RNA – and we
any methylation differences between
combine this work with other
the two. They are usually very similar,
“We then use a Spark® multimode
approaches to personalize therapies.”
and that gives us confidence that our
microplate reader to monitor the effects
screening results are faithfully
of the library compounds using a
“We have a number of drug libraries that
capturing the expected drug
CellTiter-Glo® Luminescent Cell Viability
we use to screen brain tumor cultures,
responses in patients.”
Assay (Promega). We have a Spark-Stack™ plate stacker which automates our plate
looking for tailored therapies that could be effective against individual tumors.
“When our lab was first set up in 2015,
loading, so we can just set everything up
The library that we are most interested
we were going to perform the whole
and walk away – it’s so convenient. It’s
in – which was developed by one of our
workflow manually in a 96-well plate,
also much more consistent and reliable
team members – is comprised of 200 of
but it was clear that automation would
than our previous equipment –
the most promising compounds that are
reduce our material consumption and
improving the reliability of our data –
currently being used clinically or are in
increase productivity. We invested in a
and it’s straightforward to set up and
phase III clinical trials, and we need
D300e Digital Dispenser, allowing us to
use. We have an amazing team working
about two million cells for this screen.
dispense nine dilutions of each of the
on the drug pipeline, most of them
We now have a second, more extensive
drugs from our libraries into plates prior
medical students coming through the
library that contains around 650
to drug screening. We print between 60
lab. Although the majority of them have
30
TECAN JOURNAL 1/2019
DRUG DISCOVERY
he correlation T scores were very high, proving just how reliable and consistent the platform is – it’s amazing.
The Pediatric Neuro-Oncogenomics department at the University Hospital Düsseldorf
never touched a pipette before, they are able to use the equipment after a short tutorial. David Pauck, one of our medical students, even developed the algorithm we use to generate our IC50 values, which takes the raw data straight from the reader. This has been invaluable, and
To find out more about Tecan’s D300e Digital Dispenser, visit www.tecan.com/D300e To find out more about Tecan’s
emphasizes how user friendly the
Spark multimode reader,
platform is. It’s a real team effort.”
visit www.tecan.com/spark
“The automation solutions from Tecan
To learn more about the
are amazing; they are extremely
work of the Pediatric Neuro-
consistent and help us to generate
Oncogenomics group, go to
results that we have confidence in, while
www.uniklinik-duesseldorf.de/
reducing our material consumption and
unternehmen/kliniken/klinik-fuer-
saving us countless time. This allows us to focus on our goal of translating our research into a clinical application,” concluded Daniel.
kinder-onkologie-haematologieund-klinische-immunologie/ forschungsbereiche/paedneuroonkologie
TECAN JOURNAL 1/2019
31
The Blog TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU. www.tecan.com/blog
Every lab. Every day. Empowered. Australia +61 3 9647 4100 Austria +43 62 46 89 330 Belgium +32 15 42 13 19 China +86 21 220 63 206 France +33 4 72 76 04 80 Germany +49 79 51 94 170 Italy +39 02 92 44 790 Japan +81 44 556 73 11 Netherlands +31 18 34 48 17 4 Nordic +46 8 750 39 40 Singapore +65 644 41 886 Spain +34 93 595 25 31 Switzerland +41 44 922 89 22 UK +44 118 9300 300 USA +1 919 361 5200 Other countries +41 44 922 81 11
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 © 2019 Tecan Trading AG, Switzerland, all rights reserved.
www.tecan.com
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