Tecan Journal
Edition 2/2017
Life Sciences, Diagnostics and Partnering
Fishing for genetic information
Traditional medicine meets modern analytics
Perfect synergy for molecular diagnostics
Luciferase comes to the devil’s rescue
Pages 8-9
Pages 14-15
Pages 26-27
Pages 28-29
CEO WELCOME
Welcome Dear Reader, Here at Tecan, we are committed to providing our customers with best-in-class laboratory automation solutions that combine cutting-edge performance with exceptional reliability. This is what the Tecan brand is known for in the marketplace. It is also why leading diagnostics and life science instrument providers trust Tecan Partnering to help them build some of the most successful products on the market. Our recently announced agreement with IVD specialist DiaSorin is a good example of the trust our partners place in us. Our FluentÂŽ Laboratory Automation Solution will be used in combination with the DiaSorin LiaisonÂŽ MDX PCR instrument to provide customers with a complete sample-to-result system for molecular diagnostics. The Fluent platform is already popular for drug discovery and clinical research applications (pages 20-23), and this will be the first time it features in an OEM project. This issue of the Tecan Journal contains several more examples of how Tecan Partnering can help customers to accelerate and simplify instrument development, offering everything from complete automation of a proprietary technology (pages 26-27) to supply of liquid handling robotics for easy integration into a customer-developed solution (pages 10-11). These OEM development capabilities perfectly complement our portfolio of life sciences automation solutions, and it is thanks to this in-depth application knowledge and broad expertise that we remain the partner of choice for research laboratories and leading instrument manufacturers around the world. Dr David Martyr CEO
2
TECAN JOURNAL 2/2017
CONTENTS
Contents 2
CEO Welcome
4 - 7 The time is now – automation for the academic laboratory
8 - 9
Fishing for genetic information
10 - 11 OEM instrument design offers automated patch clamping solution
12 - 13 8-9 Fishing for genetic information
Working towards cancer-free childhoods
14 - 15 Traditional medicine meets modern analytics 16 - 17 Accelerating R&D through collaboration and automation
18 - 19
Taking the guesswork out of drug development
20 - 21 What is normal? Understanding the vaginal microbiome
22 - 23
Flexibility for speed
24 - 25 Faster processing of samples in steroid hormone testing
26 - 27 Perfect synergy for molecular diagnostics 28 - 29 Luciferase comes to the devil’s rescue 30 - 31
30 - 31 Expanding the experimental space
Expanding the experimental space
TECAN JOURNAL 2/2017
3
BLOG
The time is now – automation for the academic laboratory
Pluripotent stem cell
The Blog 4
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 2/2017
BLOG
‘There is no time like the present’
think about the day-to-day
perfectly applies to the topic we are
challenges you face, and
about to discuss: integrating automated
what it takes to keep
solutions into the workflows of
everything operating
academic laboratories. The oft-repeated
smoothly and efficiently,
and stale arguments made against the
producing high quality data.
need for automation in an academic
If you break down your typical
setting are dated and, in most cases, no
processes and protocols into individual
longer relevant. There are myriad
steps, it will become increasingly clear
advantages to introducing automated
where there are gaps and potential
liquid handling, detection and control
sources of error, and where you can gain
systems to any lab – whether it’s a
the most from improving accuracy,
single, dedicated instrument designed
throughput and efficiency. Whether your
to carry out a specific labor-intensive
laboratory is focused on genomics or
task, a modular system that can be
proteomics research, doing cell-based
reconfigured and expanded as needed,
assays, ELISAs or next generation
A point well taken. Lab technicians,
or a complete workstation with
sequencing, automation can help you
postdocs and graduate students are all
integrated components to automate
to meet your individual needs.
quite capable of manually pipetting
throughput, but it also improves pipetting accuracy and precision – the two pillars of data reliability. “I don’t need robotics. I have enough staff to do everything manually.”
liquids, plating samples, feeding cell
your entire workflow. We will explore these options and discuss what your lab
“I don’t do the kind of high
cultures and performing assays.
could gain in terms of precision,
throughput experiments that can
However, the major problem with any
reliability, data quality, reproducibility
benefit from automation.”
manual activity is the risk of human
and productivity, as well as try to dispel
Automation doesn’t just process
some of the myths that many people still use to convince themselves that automation has no place in academia. The elephant in the room – cost The first argument is that ‘automated instruments are too costly – I just can’t afford them.’ Research budgets are tighter than ever, and every expense must have proven value. Not only can an experienced laboratory automation provider understand what you do and don’t need, they can also answer your questions and concerns – helping you to explore instrument and system options that best serve your current requirements – offer the flexibility to adapt to meet future needs, and meet your budgetary constraints. With this in mind, Tecan has put together a special pricing program to empower academia, making it more affordable than ever to automate your workflow. Why fix something that isn’t broken?
samples faster. Whether you are working with 100 samples or 1,000, accuracy and precision are essential for obtaining consistently high quality results. Reproducibility is the cornerstone of
error. This can be frustrating and lead to unexplained results, but it can also be costly in terms of time and resources if you have to repeat experiments. Consider too: • People need to take breaks during
scientific experimentation, and an
the day, and vacations (they also
inability to reproduce results has,
daydream and have lapses in
unfortunately, led to several recent
attention)
retractions of published papers in high profile journals. Standardization of
• People get sick
experimental methods to the extent
• People might leave for new positions
possible with automation minimizes
once they have mastered a protocol,
variability between runs, contributing to
requiring the training of new staff
more reproducible results that you can present with greater confidence. Inaccurate pipetting can have damaging effects on results and data quality across
• People get bored of doing tedious, repetitive tasks, and they are at risk of repetitive stress injuries • The accuracy and precision of manual
the workflow, whether you’re setting up
pipetting varies more than that of a
screening protocols or performing serial
robotic liquid handling platform (and
dilutions. Poor pipetting performance
you cannot simply recalibrate your
can have major consequences for highly
technicians periodically!)
sensitive applications such as NGS library preparation – with its many tedious and error-prone steps – and can result in time-consuming and expensive
You may be convinced that your
resequencing.1 Automation of library
laboratory is running well, and cannot
preparation minimizes errors, reduces
really benefit from automation. But
hands-on time and enables higher
www.tecan.com/blog TECAN JOURNAL 2/2017
5
BLOG
control, especially for extended time course experiments.3 Stem cells and induced pluripotent stem cells are especially sensitive to changes in temperature, humidity and other environmental conditions. A fully automated workflow with an integrated incubator, robotic liquid handling to minimize the risk of contamination, and a plate reader with built-in environmental controls can improve outcomes and data quality. This will not only increase your lab’s productivity, it can also save you from the cost and inconvenience of repeated experiments, wasted samples and reagents, and missed publication or presentation dates. Consider too that, if you can use walkaway automation to free up the valuable time of your graduate students and postdocs, they can be using their
Mutant stem cell
energy and expertise in more A study published in the Journal of Laboratory Automation tested the effects of a miscalibrated liquid handling system on assay performance.2 The
purposeful ways. “Automated systems are too complicated to learn and operate.”
study involved two types of in vivo
A decade ago, that sentiment may have
biochemical assays, and demonstrated
been valid, but it is no longer the case.
that small changes in accuracy did affect
You can now choose from a selection of
assay performance. While there was no
ready-to-run, standalone instruments
appreciable change in signal-to-
and fully integrated turnkey systems,
background ratio or assay variability
many of which are available with built-in
(Z-factor), inaccurate dispensing had a
training videos, user guides and
measurable effect on compound
application protocols, as well as training,
potency results, and was ‘especially
technical support and ongoing
drastic for lower potency compounds’.
maintenance options. These intuitive,
The data showed that lower potency
automated workstations require little
compounds could easily be missed, and
more than loading of the materials,
that the assay loses resolution for high
telling the system what you want it to do
potency compounds that have similar
and pushing ‘start’, with preprogrammed
IC50 values.
software scripts to take the hassle out of setting up experiments.
Working with a living system When performing live cell assays, keeping your cells healthy and alive until the assay is completed requires
The Blog 6
“Automated systems are not flexible enough – I don’t know what my needs will be in the future.”
substantial hands-on time, careful
The open architecture platforms such as
monitoring and robust environmental
the Fluent® and Freedom EVO® combine
TRENDS, NEWS, STORIES AND MUCH MORE! FROM THE EXPERTS TO YOU.
TECAN JOURNAL 2/2017
BLOG
exceptional ease of use with the ability to customize protocols as needed. No matter how you define ‘flexible’, consider the following: • Workstations can accommodate one or more liquid handling or gripper arms, with various pipetting heads to suit a broad range of applications and dispensing volumes
References 1) Sitarska, A. Precision and accuracy – two pillars of data reliability. Tecan blog. www.tecan.com/blog/precision-and-accuracy-twopillars-of-data-reliability 2) Hentz, NG, Knaide, TR. Effect of liquid-handling on assay performance. J Lab Automation, 2014, 19(2), 153-162. 3) Oberdanner, C. Great expectations for your live-cell assays? Here’s how to keep cells performing even when you’re asleep. Tecan blog. www.tecan.com/blog/great-expectations-for-yourlive-cell-assays-heres-how-to-keep-cells-performingeven-when-youre-asleep
• Platforms can be fully integrated with your choice of peripheral devices, including incubators, plate readers, shakers, mixers, etc. • Modular instrument designs make it easy to reconfigure workstations as research needs change, adding, removing or upgrading individual components • Systems can sit on a lab bench or be freestanding to take advantage of the space below the workdeck, including robotic access to below-deck instruments, devices or storage From individual components to fully integrated solutions, there are now a variety of options to automate the workflows of academic laboratories. Experienced laboratory automation providers understand the specific needs of the sector, and can help you to select the instruments, software and system configurations best suited to your laboratory. And, with our new pricing structure making these industry-proven automated solutions affordable for academia, the time is now. What are you waiting for?
Stem cells
www.tecan.com/blog TECAN JOURNAL 2/2017
7
MARINE BIOLOGY
Fishing for genetic information Canada’s Molecular Genetics Laboratory at the Pacific Biological Station uses DNA analysis to identify and track salmon from different hatcheries. Automated NGS has enabled the laboratory to introduce parentage-based tagging, a cost-effective alternative to the coded wire tag system.
The Pacific Biological Station in
produced salmon is incomplete, as not
the Pacific Salmon Treaty between
Nanaimo, British Columbia, is part of a
all fish are tagged. An alternative, more
Canada and the United States. To
network of nine major scientific facilities
cost-effective approach is parentage-
adhere to the terms of the treaty, it’s
operated by Fisheries and Oceans
based tagging (PBT), which involves
important to determine the origins of
Canada, the government agency
genotyping of hatchery broodstock.
both Chinook and Coho salmon in
responsible for the management of
Establishing a database of broodstock
fisheries. Historically, this has been
Pacific salmon fisheries. The oldest
genotypes from each hatchery allows
done using CWTs, but can now be
fisheries research center on the Pacific
the offspring to be non-invasively
achieved with PBT technologies. PBT
coast, it is home to the Molecular
sampled to identify the hatchery and
gives a more refined estimate of stock
Genetics Laboratory (MGL), which is
brood year according to parentage.
compositions than using CWTs, helping
responsible for determining Pacific
Once complete, all juvenile fish are
to determine the contributions of each
salmon population structure and
effectively tagged and can be traced to
hatchery to commercial and
estimating stock compositions in
a specific hatchery.
recreational fisheries. If there is concern about conservation of a particular
mixed-stock salmon fisheries. The abundance of Chinook and Coho salmon in British Columbia has been enhanced by production from hatcheries, with a portion of the juveniles marked with coded wired tags (CWTs) prior to release. If an individual containing a CWT is recovered from a
I can foresee that [NGS] is going to become the way of the future in a lot of applications.
stock, knowing the timing and location of the stock enables the closure of specific areas at appropriate times.” Terry continued: “Previously, stock composition was primarily determined by microsatellite analysis, and this technique is still a major workhorse in our lab, particularly for those
fishery, its age and hatchery of release can be determined. However, tagging
The MGL has been using DNA analysis
applications where we need an answer
with CWTs can be expensive, the tag
since the early 1990s, as research
within 24 hours. During the fishing
cannot be retrieved without killing the
scientist Terry Beacham explained: “The
season, we use microsatellites to
fish, and identification of hatchery-
catch of Pacific salmon is governed by
analyze several thousand Pacific salmon in just a few weeks, estimating the stock composition to help decide when a fishery should be opened or closed. More recently, we have implemented automated NGS protocols, allowing hundreds of single nucleotide polymorphisms (SNPs) to be costeffectively analyzed. When we first considered using SNPs for genetic stock identification, the number of SNPs we would need to run to generate results equivalent to that of microsatellite analysis – and the cost and throughput of the platforms available at the time – meant it simply wasn’t viable. With the advent of
The Pacific Biological Station team 8
TECAN JOURNAL 2/2017
MARINE BIOLOGY
NGS protocols, we invested in an Ion
sequencing verified, flexible protocols
Proton™ System – enabling us to
for Ion AgriSeq library preparation,
cost-effectively analyze several hundred
developed in collaboration with Thermo
SNPs at one time – and have developed
Fisher Scientific. One of our technicians
methods to monitor Chinook and Coho,
attended a Freedom EVOware® training
the main species of salmon produced in
course, and the knowledge she gained
hatcheries in British Columbia.”
from this allows her to write many scripts for users from different groups,
To take full advantage of the
which is a real benefit. We’ve been
throughput capabilities of the Ion
running the systems for about two
Proton System, and to provide
years now, developing the workflow
additional flexibility in the lab, the MGL
and optimizing the scripts for our
invested in two Freedom EVO® 100
applications, and can now run four chips
workstations. “Typically, we receive 100
a week – representing over 1,500 fish –
adipose fin punches attached to a
which is a fairly good throughput.
Whatman filter paper. DNA is extracted
While NGS is quite a new technology
from these punches either manually
in fisheries management, I can foresee
using the Chelex method or automated
that it is going to become the way of
on a BioSprint instrument, ready for
the future in a lot of applications,”
Applied Biosystems AgriSeq™ library
concluded Terry.
Fisheries and Oceans Canada is responsible for monitoring the Pacific salmon population
preparation. Initially, DNA is amplified and normalized to a concentration of
All Tecan products mentioned are for research
40 ng/µl using a Freedom EVO
use only. Not for use in clinical diagnostics.
platform equipped with an Air LiHa™,
Punches are taken from the adipose fin
before adding the primer panel to the 96-well plate using the second workstation’s MultiChannel Arm™ 96. Currently, we look at between 300 and 350 amplicons depending on the species, but hope to extend this to 500 amplicons or more. After digestion, a set of 384 barcodes is ligated to the amplified products, and the pooled libraries are transferred to an Ion Chef™. This generates two sequencing-ready chips for the Ion Proton System, from which we determine the genotype of
To find out more about Tecan’s genomics solutions, visit www.tecan.com/genomics To learn more about the Fisheries and Oceans Canada Pacific
the fish.”
Biological Station, go to
“We chose Tecan because it is a well-
facilities-installations/pbs-sbp/
known brand with a good reputation and
index-eng.html
www.pac.dfo-mpo.gc.ca/science/
Samples are mounted on filter paper and sent to the laboratory
quality products. There is also the added advantage that the company offers
TECAN JOURNAL 2/2017
9
PATCH CLAMPING
OEM instrument design offers automated patch clamping solution Ion channels regulate many
Sophion Bioscience, in Ballerup,
with a cell’s membrane. A small amount
physiological processes, as
Denmark, was established in 2000, and
of suction ruptures the membrane to
among its aims was a desire to ‘take the
bring the cytoplasm and pipette
voodoo out of patch clamping’. Since
solution in direct contact, enabling the
its start-up days as a spin-out from
electrode to measure currents between
NeuroSearch, its team of dedicated
10-12 and 10-9 A passing through the ion
drugs. Patch clamping remains
electrophysiologists has expanded to a
channels.”
the gold standard assay for
company of around 50 employees
investigating ion channels, but
designing automated patch clamping
“Traditional, manual patch clamping
solutions. Rasmus Bjørn Jacobsen, head
requires a high level of skill, and it can
of the applications development group,
take a year’s training to become
explained: “Patch clamping is a direct
proficient. Even then, throughput is
measurement technique used to
resulting in ion channels
limited to five to ten assays per day,
investigate the passage of ions across
and each practitioner has their own
remaining poorly understood.
cell ion channels. The whole-cell
equipment, which raises concerns
method uses an electrode-containing
about reproducibility. Alternatives to
glass pipette to create a gigaohm seal
patch clamping involving fluorescent
well as playing a role in many diseases, making them a target for 20 percent of registered
the manual technique requires patience, and extensive training, and has a low throughput,
The Q-Patch offers the accuracy of manual patch clamping with increased throughput and reproducibility
10
TECAN JOURNAL 2/2017
PATCH CLAMPING
dyes have a much higher throughput, but suffer from low sensitivity and accuracy. With such obstacles to overcome, it made sense to design an automated solution.” In 2004, the company launched QPatch® – a fully automated patch clamping system – into the market. Göran Mattsson, global product manager for QPatch, said: “The whole assay process is handled by the instrument, from cells to compounds to consumables. The user simply presses ‘Start’ and the experiment runs for up to five hours. This unattended operation is an essential requirement of many pharma companies, who also demand high quality standards and reproducibility. Three quarters of the top 20 pharma companies worldwide are now using a QPatch – or other Sophion products – for drug discovery and safety testing, and it has become the benchmark for
The Xantus can switch between multichannel and single channel pipetting
cardiac safety.” “The QPatch brings together the
design; the power and the cables
accuracy of manual patch clamping
connect to the back end, and everything
with the high throughput of alternative
else – including the pump – resides in
methods, and can make up to 48
the robotic arm. Crucially, the Xantus
recordings in 30 minutes. Training a
can switch between multichannel and
technician on the instrument takes just
single channel pipetting, with the option
two days, and the assay process does
to cherry-pick samples, and we can
not require expert oversight. One
upgrade the system when a customer
person can prepare the cells, another
requires increased throughput.”
can start the process, and someone else can analyze the results. The built-in
“It’s been great to work with Sias’
security is also an added benefit; the
in-house engineering team and we
QPatch connects to an Oracle database,
collaborated quite closely to develop
which logs all data and is FDA 21 CFR
the QPatch. Not only did they help with
Part 11 compliant. The data cannot be
the integration of the Xantus robot,
deleted or overwritten, providing
they also designed a special titanium
reassurance to pharma companies that
oxide coating for the pipette probes to
they can trust the data recorded
help prevent compounds sticking to
anywhere in the world.”
them. Sias set the bar for customer
P artnering with Tecan allows us to deliver the complete service package, not just a high quality instrument.
service support by providing valuable Jørgen Due, head of service, continued:
training to our engineers, and this
“Patch clamping requires the utmost
standard has continued under Tecan. In
precision and accuracy, and is extremely
the future, if we are bringing another
sensitive to electrical noise given the
product to market, then I’m sure our
tiny currents we are measuring. When
strong relationship is set to continue.
designing the QPatch, we looked at
Partnering with Tecan allows us to
what OEM liquid handling solutions were
deliver the complete service package,
available on the market, and we came
not just a high quality instrument,”
across the Sias® Xantus® robotic
Jørgen concluded.
pipetting platform from Tecan. The Xantus fulfilled our specific needs, and
All Tecan products mentioned are for research
we appreciated its self-contained
use only. Not for use in clinical diagnostics.
To find out more about partnering with Tecan, visit www.tecan.com/partnering To learn more about Sophion Bioscience, visit www.sophion.com
TECAN JOURNAL 2/2017
11
DRUG DEVELOPMENT
Working towards cancer-free childhoods Developing cancer drugs for clinical trials involves not only identifying and evaluating suitable agents, but also observing how they interact with the cocktail of other drugs in a cancer treatment regime. For the Telethon Kids Cancer Centre in Perth, Western Australia, increasing throughput and reducing assay volumes are essential to save money and time in the race to beat cancer. The Telethon Kids Cancer Centre (TKCC)
tumor cells for drug screening and
the D300e, it was a lightbulb moment.
brings together a dedicated group of
preclinical testing such as drug
This instrument has transformed the
researchers and clinicians from across
sensitivity assays, diluting drugs from
activities of our lab. Previously, working
the globe to work collaboratively
high to low concentrations to identify
together with a colleague, you could
towards developing more effective cures
effective doses. What’s important for
generate a small number of plates
to treat childhood cancer. Brain tumors
us is looking at how different drugs
working solidly for three to four hours.
are the most common form of solid
interact with each other; if we were to
With automation, our throughput went
tumor in children, affecting 200 children
put a new drug into an existing
through the roof and increased by four
in Australia each year, and many more
protocol, would the drug interfere with
or five times; we are now able to
worldwide. Within the TKCC, the Brain
the current treatment?”
generate faster and more consistent
Tumour Research Program (BTRP) –
results. Working manually, we would
headed up by Dr Nick Gottardo and
Three years ago, the team looked to
often see a lot more inter-experiment
Dr Raelene Endersby – is striving to
develop the automation in its workflow,
variability than we now do with the
improve patient survival rates and
to help carry out the thousands of
D300e, where all of the data overlays
quality of life, through basic and
necessary drug tests. Raelene continued:
beautifully. The error bars are much
preclinical research, with a key focus on
“We were doing lots of dilutions, and
smaller compared with our previous
providing the necessary evidence to
serial dilutions, which we were carrying
data, and the increased reliability means
help a therapy transition to clinical trials.
out either manually, or using a much
that we don’t need to repeat the
slower robot. When we found out about
experiment as many times.”
The TKCC is home to a diverse range of researchers from academia and industry, as well as clinical oncologists, neurosurgeons, radiologists, chemists, pharmacologists and bioinformaticians. It has a close connection with oncologists at the Princess Margaret Hospital – the only children’s hospital in the state – and this relationship is key to defining the direction of its research, based on the lack of treatment options currently available to patients. The aim is to uncover more targeted, less DNA-damaging treatments to tackle the tumors. Raelene explained: “My lab is interested in looking for agents that sensitize cancer cells to conventional treatments, such as chemo- and radiation therapy, and then evaluating them for safety and efficacy. We generate in vitro and in vivo models of pediatric brain cancer using surgical specimens or genetically modified mice, and then use those 12
TECAN JOURNAL 2/2017
Left to right: BTRP team members Tracy Seymour, Hetal Dholaria, Stacey Fazio, Brooke Strowger, Raelene Endersby, Mathew Ancliffe, Hilary Hii and Jacqueline Whitehouse
DRUG DEVELOPMENT
Dr Jacqueline Whitehouse, senior
“It’s very easy and quick to set up; you
scientist in the BTRP, agreed: “What
just copy and paste your data from
would take hours on our previous liquid
Excel®. It’s been simple to train people
handling robot, now takes minutes on
and there’s not much people can do to
the Tecan system. The fact that we can
break it; the leukemia group within the
use small picoliter volumes also saves
TKCC is also using our system and it’s a
money, as some of the novel anticancer
good instrument to share with people
drugs are quite expensive and hard to
knowing that it will come back in one
synthesize. Using manual pipetting or
piece. We’ve used it pretty heavily for
our previous robot, we could only work
the past two years and, since there are
with a minimum volume of about five
not many moving parts, it’s been great,”
microliters. We would be wasting a lot of
Jacqueline concluded.
W ith automation, our throughput went through the roof… we are now able to generate faster and more consistent results.
drugs just by the nature of the pipettors and their accuracy, whereas we can now
All Tecan products mentioned are for research
dispense picoliter volumes consistently.
use only. Not for use in clinical diagnostics.
We were skeptical at first, but were pleasantly surprised when we were still seeing effects of our drugs on cells using such small volumes. The other advantage is its size, it’s so small that it can easily fit inside a biosafety cabinet. This is important for both our precious cultured cells – minimizing the risk of contamination – and for the user, protecting them from exposure to potentially hazardous or toxic novel chemotherapeutics.”
To find out more about Tecan’s liquid handling and automation solutions, visit www.tecan.com/mdx To learn more about Telethon Kids, visit www.telethonkids.org.au
TECAN JOURNAL 2/2017
13
DRUG DISCOVERY
Traditional medicine meets modern analytics Chinese medicine combines herbal remedies with acupuncture, massage, exercise and diet to provide alternative therapies for a wide range of conditions. Despite drawing on over 2,500 years of traditional knowledge, little is known about the mode of action of these herbal medicines. Researchers at Zhejiang University’s College of Pharmaceutical Sciences are looking to address this, using modern laboratory techniques to identify the numerous active pharmaceutical ingredients and synergistic effects that contribute to their efficacy.
Many modern medicines – from aspirin
The College of Pharmaceutical Sciences
or combination of components is
and codeine to the chemotherapy drug
at Zhejiang University is working to
responsible for a given biological effect.
paclitaxel (Taxol®) – are derived from
uncover the secrets of traditional Chinese
We have overcome this by isolating
natural sources. In fact, it is estimated
medicines, analyzing some of the 300+
crude extracts, then separating these into
that up to 50 percent of clinically
herbs used in these preparations.
individual components or fractions based
approved drugs since 1981 have been
Professor Yi Wang explained: “Many
on size, hydrophilicity, etc. Using this
either directly or indirectly derived from
traditional medicines used in China to
approach, we have built up a component
natural products.1 As small molecule
treat illnesses, injuries and infections have
library of over 30,000 fractions over the
pharmaceutical discovery pipelines have
over 1,000 years of history, but few have
last decade, which we can screen against
begun to falter, many companies are
been studied to identify the active
cells or specific molecular targets.”
looking towards ethnopharmacological
components. The main challenge we face
products – or traditional medicines as
is that the chemical composition of most
“For this work – which formed part of my
they are more commonly known – as a
herb extracts is extremely complex.
PhD research – we used an Infinite® F200
source of novel active pharmaceutical
Chromatographic separation of a single
multimode microplate reader to develop
ingredients (APIs). The problem with this
extract can yield hundreds or even
a large number of screening assays,
approach is that few traditional medicines
thousands of compounds, making it
including several cytotoxicity assays to
have been studied to identify their APIs.
difficult to identify which component
identify antitumor and antioxidative compounds. At that time, we chose the Infinite reader for its ease of use and automated injector module, which allowed us to quickly screen numerous components. This system has become the workhorse of the lab – it still works just as reliably today – but, on the recommendation of our local Tecan representative, we have since acquired a monochromator-based Infinite M1000 to help us perform more sensitive analyses and establish new applications.” Professor Wang continued: “The free wavelength selection possible with the Infinite M1000 has helped us to design several fluorescent probes that can report on the up- or down-regulation of the activity of certain enzymes. For example, we have created a fluorescent probe for sirtuin 1 activity – a protein thought to play a role in intracellular regulatory processes – that could not
The group has built up a large component library to simplify screening 14
TECAN JOURNAL 2/2017
previously be monitored
DRUG DISCOVERY
T his is such a powerful tool for drug screening, allowing very accurate dispensing of extremely low volumes of compounds.
spectroscopically. We used this probe to
“The main advantage of the D300 is that
1) Newman, DJ, Cragg, GM. Natural products
test a panel of around 200 candidate
it can automatically set up synergy
as sources of new drugs over the 30 years
compounds for sirtuin 1 activation or
experiments, calculating and accurately
from 1981 to 2010. J Nat Prod, 2012, 75(3),
inhibition, and actually found 13 active
dispensing the required volume of each
311-35.
compounds – which is a very high hit rate
compound in a randomized layout to
All Tecan products mentioned are for research
compared to random screening.”
produce clean dose-response curves.
use only. Not for use in clinical diagnostics.
This is extremely difficult to perform by “Our work has taught us that many
hand, as there are accuracy and
Chinese medicines appear to rely on
reproducibility issues when performing
synergistic effects between multiple
the necessary dilutions to achieve
components, rather than a single API.
nanomolar concentrations of individual
Although there are some very potent
components. The D300 eliminates these
compounds, in many cases, the effects of
problems, especially for very sensitive
a single isolated component is weaker
cell viability assays, producing beautiful
than a comparable, internationally-
dose-dependence curves and helping us
University’s College of
registered small molecule drug. In
to select the best ratios of compounds.”
Pharmaceutical Sciences, go to
contrast, several compounds working in
To find out more about Tecan’s drug discovery solutions, visit www.tecan.com/drug_discovery To learn more about Zhejiang
www.cps.zju.edu.cn
combination can provide a much more powerful cellular response. Investigating these drug synergies can be very difficult to achieve manually, as pipetting small volumes of multiple drugs in complex microplate patterns is difficult to perform accurately and mistake-free. However, during a year working as a visiting scholar at Massachusetts General Hospital in Boston, USA, I was lucky enough to use a D300 Digital Dispenser for cancer drug screening. This is such a powerful tool for drug screening – allowing very accurate dispensing of extremely low volumes of compounds – and so, when I returned to China, it was the first piece of equipment I wanted to purchase.”
Members of the traditional medicines research group (left to right): Hao Li, Hao Liu, Yu Zhao, Shujing Zhang and Professor Yi Wang TECAN JOURNAL 2/2017
15
DRUG DEVELOPMENT
Accelerating R&D through collaboration and automation Discovering and developing
SMALTIS, established in 2014 by Dr
interactions by taking regular images
new antimicrobial drugs to
Sophie Guénard and Dr Cédric Muller, is
over the course of five hours. When we
a CRO specializing in microbiology,
started performing aggregation assays,
molecular and cellular biology. It offers
our customer gave us a specific
services to private and public research
operating procedure which was
laboratories, assisting with R&D
designed for a high content imaging
to new compounds. A range of
projects by providing customized
platform. As these systems are
tests are needed to determine
bacterial strains, evaluating the activity
expensive, we looked at alternative
the efficacy of potential
of antimicrobial compounds and
approaches, and developed a manual
analyzing microbial samples. The
set-up: we incubated the plate on a
company was born out of a shared
rotating incubator and removed it every
research background, as Cédric
hour for analysis, then returned the plate
explained: “Sophie and I studied
to the incubator. Although this worked
company in Besançon,
microbiology at the Patrick Plésiat labs,
well, it was very labor intensive.”
France, test automation
seeking to understand the antibiotic
has dramatically improved
resistance mechanism exhibited by
“We then came across a Spark®
Pseudomonas aeruginosa, the primary
multimode microplate reader in one of
cause of death in cystic fibrosis. We
the research laboratories of Jean-Minjoz
were fortunate to work with a big
Hospital that we collaborate with. We
pharma company to develop a specific
developed an aggregation assay
antibody vaccine and, thanks to the
protocol on the hospital’s instrument,
project, we got the idea of creating a
and a member of the Tecan team helped
company dedicated to helping
us to perform some validation
laboratories accelerate their
experiments to confirm that we could
microbiology R&D. We perform
transfer our technology and methods
minimum inhibitory concentration (MIC)
onto the system. We then went ahead
studies and time-kill experiments,
and purchased our own Spark reader,
characterizing how the test compounds
allowing us to perform the same
act on the target bacteria, and how the
aggregation assay far more cost-
bacteria in turn adapt to the drug,
effectively than using a high content
helping our clients to improve the
imaging platform.”
tackle antibiotic resistance requires an understanding of how bacteria respond and adapt
drugs, such as aggregation and adhesion/invasion assays. For SMALTIS, a biotechnology
throughput and data collection, freeing up research hours to concentrate on developing new experiments.
efficacy of their compounds.” Cédric continued: “The experiment now Aggregation and adhesion/invasion
runs without the need for supervision,
assays are among the many services
with the added benefit that the Spark
SMALTIS offers. In order to colonize and
produces growth curves of the strains
initiate an infection, bacteria adhere to
over the course of the assay. Automation
host cells via the fimbriae (hair-like
has freed up our time to develop other
structures) that interact with specific
experiments, for example, we are
host cell receptors. Aggregation assays
currently developing a method to
are used to assess the ability of a
evaluate luminescent bacterial strains,
compound to inhibit adhesion, by
created by inserting a copy of the
interacting with the fimbriae to cause
luxCDABE operon – from Photorhabdus
aggregation. Cédric said: “Our basic
luminescens – into their DNA.”
assay monitors drug-bacteria
16
TECAN JOURNAL 2/2017
DRUG DEVELOPMENT
W hat took one person three days, now takes an hour and a half.
“We also use the Spark to carry out
adhesion/invasion capabilities. Prior to
adhesion/invasion assays to determine
having the Spark, we had to take four
the capacity of bacterial strains to
to six images to cover each 96-well
attach to or invade eukaryotic cells.
plate, and analyze the colours with
The cells are infected with a bacterial
a microscope. This is now done
strain, then after a series of washes,
automatically, and what took one
the eukaryotic cells are stained with
person three days, now takes an hour
DAPI and the bacteria with specific
and a half. We are very happy with the
antibodies conjugated to an AlexaFluor®
Spark; it easily adapts to each assay
dye. We measure the blue and green
set-up, and its high throughput and
fluorescence, from the eukaryotic cells
automation has enabled us to
All Tecan products mentioned are for research
and bacteria respectively, to determine
continually improve our service for
use only. Not for use in clinical diagnostics.
a ratio reflecting the strain’s
our clients,” Cédric concluded.
To find out more about Tecan’s Spark reader, visit www.tecan.com/spark To learn more about SMALTIS, visit www.smaltis.fr
Left to right: SMALTIS team members Julie Challant, Sophie Guénard, Marjorie Robert-Nicoud, Cédric Muller (back), Elise Ponçot and Charlotte Richardot-Saura TECAN JOURNAL 2/2017
17
CELL BIOLOGY
Taking the guesswork out of drug development California-based company zPREDICTA™ has created a novel technology that reconstructs physiologically-relevant organ-specific human microenvironments that help eliminate the guesswork from drug development. Meaningful drug discovery studies involve complex experiments that are not feasible to perform manually. Automation is the answer, improving accuracy, saving time and reducing the amount of compound used.
A major problem in drug discovery and development is that a high proportion of the drugs reaching clinical trials – around 95 percent – ultimately fail due to a lack of efficacy. In part, this is because non-physiological 2D cell culture and mouse models frequently
used in preclinical drug discovery have
on the basis of a poor performance in
limited effectiveness as a representation
mouse models might actually work very
of human diseases. This can result in a
well in people. Based on the principle
poor correlation with the response
that cures for human diseases require a
observed in clinical trials; drugs
human-specific discovery and testing
demonstrating promising results in
environment, zPREDICTA, based in San
preclinical studies in physiologically-
Jose, USA, developed a novel 3D
irrelevant models may prove ineffective,
technology for reconstruction of a
while compounds that were deprioritized
physiologically-relevant, organ-specific human microenvironment, the Reconstructed Organ (r-Organ) platform, which mimics the native architecture of human tissues in an organ- and diseasespecific manner. As a result, there is a high correlation between preclinical studies and clinical response. The r-Organ platform lies at the core of zPREDICTA’s goal to integrate different technologies and automate the various steps in drug development and establish a seamless workflow, from the initial drug screening process through to preclinical and clinical studies. Dr Julia Kirshner, founder and CEO, explained: “Our research involves three-dimensional organ reconstruction within an organspecific extracellular matrix, followed by cell culture under human disease conditions. As this process is very close to what happens in a person, when the drugs are applied to this platform they behave in the same way as they do in a patient, resulting in a high correlation with clinical response.” Julia continued: “The r-Organ platform is quite versatile – it can handle a wide range of drug types, including small molecules, antibodies, antibody drug conjugates, and CAR-T cells – and the
Dr Niyati Jhaveri uses the D300e for drug development studies
18
TECAN JOURNAL 2/2017
technology is already starting to be used
CELL BIOLOGY
his [the D300e] T has contributed to a four- to five-fold improvement in accuracy.
The intuitive D300e software makes it easy to set up an experiment
in the pharma industry. We currently offer a fully validated Reconstructed Bone Marrow (r-Bone) for the study of
Set-up time
leukemia and multiple myeloma, and are
(including calculations and making dilutions)
establishing a lung system for the investigation of non-small cell lung
Treatment time (60 plates)
cancer. We also have other tissues at
(single/combination treatment and vehicle backfill)
different stages of development and
Time savings
validation, including brain, a lymph node for the study of lymphomas, and mammary gland tissue for breast cancer. These physiologically-relevant tissues are ideal for drug development studies.” “When you are performing drug testing studies, multiple steps are involved – dilution, dose-response, combination treatment – to acquire data at different doses and various time points, and the process really needs to be automated. For instance, some larger scale projects have involved a couple of dozen patients, with 240 drugs and a seven-point dose-response curve. That’s anything from a couple of thousand wells through to 10,000 for the entire study, which is a huge undertaking manually. It is critical that there are no transfer errors, but it is very difficult to manually track every well to ensure that the contents are correct. You also need to take variation across the plate – for example, due to evaporation – into consideration by randomization of the well contents. Manual processing is just not feasible. It is incredibly tedious and extremely difficult to do without making mistakes, and that’s why we invested in a D300e Digital Dispenser from Tecan.” “We’ve had the D300e for about 18 months now, and find it very user friendly
Amount of compound needed Compound usage Hazardous chemical waste
Manual
D300e
3.5 hours
15 minutes
26 hours
2.5 hours
10× (3.3 FTE days) 68 μl
80 % reduction 20 ml
Waste reduction Accuracy (standard deviation)
13.6 μl
7 μl >2000×
11.4
Accuracy improvement
2.6 4-5×
Implementation of the D300e has led to a significant improvement in overall performance
and easy to operate. Automation has
“I really enjoy using the D300e and it’s
made a tremendous difference to our
a pleasure working with Tecan’s
workflows. Manual errors, such as missing
representatives, engineers and scientists.
wells or pipetting into the wrong well,
The team is very professional and we
have been eliminated, and we can set the
have established a great relationship,”
system to shake the plate after dispensing
Julia concluded.
to ensure even distribution and suspension of the drugs in the matrix,
All Tecan products mentioned are for research
preventing the creation of higher dose
use only. Not for use in clinical diagnostics.
pockets. This has contributed to a four- to five-fold improvement in accuracy. We save a great deal of time too. Manual set-up that took three and a half hours can be completed in just 15 minutes with the D300e, and the treatment time for 60 plates is reduced from 26 to two and a half hours. Other benefits include an 80 percent decrease in the amount of compound used, and more than a 2,000
To find out more about Tecan’s cell biology solutions, visit www.tecan.com/cellbiology
times reduction in waste, which will
To learn more about zPREDICTA,
become increasingly important as we
go to www.zpredicta.com
scale up our experiments.”
TECAN JOURNAL 2/2017
19
GENOMICS
What is normal? Understanding the vaginal microbiome Microbiome research is still in
The human microbiome – the collection
interplay of microbial populations within
its infancy, with little currently
of micro-organisms living on or within
the gastrointestinal tract. However,
our bodies – plays a vital role in our
microbiota are also thought to play a
health, from assisting in the digestion of
significant role in women’s reproductive
foodstuffs within our gut to causing or
health, and so we are looking into this
preventing certain diseases or cancers.
field too. The major problem with this is
health and the genesis of
Despite this, the composition and
that, in order to understand what is
disease. Researchers at the
maintenance of the microbiome is poorly
abnormal, first we need to know what is
Karolinska Institute are using
understood. Although some progress has
‘normal’. Because so little comprehensive
been made in this area over the last
research has been done in this area, a
decade, a majority of current research
‘normal’ baseline of the micro-organism
initiatives are directed towards the gut or
population has not been established, and
skin microbiomes, with little focus on
so we have just begun a large study to
other tissues and biofluids.
better define this.”
The Centre for Translational Microbiome
“When samples arrive in our laboratory,
Research (CTMR) – a collaboration
we begin by extracting and aliquoting
between Sweden’s Karolinska Institute,
the DNA using a Freedom EVO®
Science for Life Laboratory (SciLifeLab)
workstation. The resulting extracts are
and Ferring Pharmaceuticals – and the
quantified using a Spark® reader, then
Human Microbiome Translational
one aliquot is sent to the biobank, and a
Research Program (HMTRP) aim to
second is used for sequencing. Most of
better understand the contribution of the
our studies rely on 16S rDNA sequencing
understood about the role micro-organisms play in both maintaining our day-to-day
next generation sequencing to establish a baseline of the microbiota present in healthy individuals as a starting point for the development of new therapeutic strategies for a wide range of diseases.
H uman error is one of the greatest risks to results in any lab, and so I am a big fan of using automation to reduce hands-on time as much as possible. human microbiome to human health,
to identify the micro-organisms present,
with the goal of developing novel
which starts with DNA normalization.
therapies. Its current focus is on the
This is performed by our Fluent®
microbiomes of the gut and the female
Laboratory Automation Solution, which
reproductive organs. Maike Seifert,
uses the concentration data from the
Laboratory Engineer at CTMR, explained
Spark to calculate the exact pipetting
the center’s approach: “The gut
volumes to achieve normalization. The
microbiome was the first microbial
system then performs the library
population to be investigated, and
preparation, generating sequencing-
initiatives such as the Human Microbiome
ready samples that can be run on our
Project mean we are now beginning to
MiSeq™ platform.”
understand the composition and
20
TECAN JOURNAL 2/2017
GENOMICS
Maike continued: “We chose the Fluent system for this application because our previous liquid handling platform was not able to perform eight-channel pipetting. This meant that normalization had to be done one sample at a time, taking two and a half hours per plate and creating a major bottleneck in our workflow. In
To find out more about Tecan’s genomics solutions, visit www.tecan.com/genomics To learn more about the Centre for Translational Microbiome
comparison, our Fluent can perform the
Research, go to
same normalization in just 15 minutes,
ki.se/en/research/centre-for-
saving a huge amount of time. Another
translational-microbiome-
benefit, which we hadn’t appreciated
research-ctmr
until we began validating the system, is that the Fluent pipettes incredibly precisely; I’ve never seen an automated platform anywhere near as accurate – it’s wonderful. This will be particularly useful for whole genome sequencing, which requires extremely precise amounts of DNA from very low concentrations of starting material – as little as 0.2 µg/µl – to achieve high quality libraries.” “Using the Fluent has certainly improved the quality of our library preparations, and our laboratory staff are happier. It has also virtually eliminated the risk of pipetting errors during normalization – a task that requires a lot of focus to perform manually. Human error is one
DNA is extracted and aliquoted using a Freedom EVO workstation
of the greatest risks to results in any lab, and so I am a big fan of using automation to reduce hands-on time as much as possible, especially for high throughput studies. We currently perform one sequencing run a week, which could potentially be done manually, but using the Fluent workstation is better for both our staff and samples, and will allow us to increase this to two runs a week as our sample numbers increase,” Maike concluded. All Tecan products mentioned are for research use only. Not for use in clinical diagnostics.
TECAN JOURNAL 2/2017
21
BIOANALYSIS
Flexibility for speed Maintaining a flexible approach can be difficult in biopharmaceutical research; core laboratories must balance the ability to adapt to individual project requirements with the need for efficient, high throughput processing of ever-increasing sample numbers. Novo Nordisk’s Research Bioanalysis Department has adopted a semi-automated workflow, which combines the versatility to work across the company’s various research areas with accurate and reproducible testing of thousands of samples a day.
Originally established in the early 1920s
and pharmacodynamic profiles of drug
Principal Scientist, explained the
to manufacture insulin for the Nordic
candidates from across the company’s
department’s workflow: “We have a
market, Novo Nordisk has retained a
research pipeline. Samples from animal
total daily throughput of around
strong focus on the treatment of
or cell culture experiments are tested
35,000 tests, and use a combination of
diabetes. Today, this expertise in protein
with a panel of 100 to 200
fully and semi-automated set-ups to
and peptide technologies is also
immunoassays to gain a thorough
ensure rapid turnaround times, and to
being used to develop products for
understanding of their activities.
give us the flexibility to adapt to
hemophilia and other bleeding
Around 15 years ago, the laboratory
changing project priorities. We
disorders, as well as growth and
automated its ELISA procedures to
purchased a Fluent® Laboratory
hormone replacement therapies.
increase throughput, and improve the
Automation Solution last year to
accuracy and reproducibility of testing.
further increase our throughput, and it
Biopharmaceutical development and biomanufacturing rely on an in-depth
The sensitivity and convenience offered
understanding of the complex in vivo
by bead-based AlphaLISA®
actions of therapeutic products to
immunoassays have made it the
ensure their safety and efficacy. The
technology of choice within the
Research Bioanalysis Department,
department, and these
based at Novo Nordisk’s research
chemiluminescent assays now account
center in Måløv, Denmark, is responsible
for over 70 percent of the total
for investigating the pharmacokinetic
immunoassay workload. Johannes Fels,
is now used to perform various liquid transfers and sample dilutions as part of both our automated and semiautomated workflows. AlphaLISA is a very easy technology to run in a relatively high throughput format – there are only two reagent additions and two incubations – and so, although many of our protocols could be completely automated on the Fluent,
S taff like the Fluent platform – it is completely reliable, very fast, accurate and nice to work with. semi-automation frees up the platform for other users to perform fast assay set-up or dilutions during the incubation steps. This way, each plate only takes two or three minutes, occupying the liquid handling workstation for just a short time.” “A typical semi-automated AlphaLISA Left to right: Kirsten Jensen, Anita Svendsen and Johannes Fels with the Fluent platform
22
TECAN JOURNAL 2/2017
protocol starts with the Fluent diluting
BIOANALYSIS
samples 10-, 20- or 40-fold from mother
can perform 100-fold dilutions in a single
to daughter plates. Next, the Fluent is
step, something we have not seen on
used to transfer samples from both
any other liquid handling system.”
mother and diluted daughter plates into a 384-well assay plate containing two replicates of each source plate. After this step, the assay plates are manually transferred to a benchtop dispenser, where assay buffer containing biotinlabeled antibodies and AlphaLISA acceptor beads is added. By running both the mother and daughter plates, we can more than double the experimental window for each assay, helping us to overcome large variations in sample or analyte concentration without performing repeat testing. Plates are incubated offline for one hour, and then moved back to the benchtop dispenser for addition of streptavidin donor beads, followed by another 30 minutes incubation. Finally, the plates are transferred to a reader for
“Staff like the Fluent platform – it is
To find out more about Tecan’s Fluent, visit www.tecan.com/fluent
completely reliable, very fast, accurate and nice to work with – and everyone in
For more information on Novo
the department runs the same set of
Nordisk, go to
universal scripts, simply changing the
www.novonordisk.com
dispense volumes, dilutions, matrices and buffers between the various assay procedures. Everything we need is programmed into the user-friendly FluentControl™ interface, and I especially like the way the liquid classes are built up within the software. Our local Tecan application scientist helped me to do the initial scripting, so we were able to get the various protocols up and running as we wanted within two weeks of delivery,” Johannes concluded. All Tecan products mentioned are for research use only. Not for use in clinical diagnostics.
chemiluminescence measurements.” “Accurate and reproducible low volume dispensing is vital to achieve reliable assay performance using this very sensitive immunoassay technology, and we worked closely with the local Tecan team to design a site acceptance test that would demonstrate the suitability of the system for our needs. We were immediately able to achieve very low CVs of 2-3 % for 1 µl plasma transfers into dry 384-well assay plates, so we were confident that it would perform as expected. Another nice feature is the extended volume tip adaptor, allowing us to combine four channels of the Multiple Channel Arm to perform 96-well dispensing of up to 500 µl at a time. This makes it very versatile; you
Kirsten Henriksen (left) and Annette Larsen return samples to the hotel
TECAN JOURNAL 2/2017
23
CLINICAL BIOCHEMISTRY
Faster processing of samples in steroid hormone testing Mass spectrometry is now routinely used for clinical diagnostics around the world, but manual sample preparation and traditional liquid-liquid extraction techniques are very time consuming, requiring new approaches to help streamline laboratory workflows. The endocrinology service at Charing Cross Hospital is using high throughput automated SPE to accelerate mass spec sample prep, improving turnaround times and freeing up staff time to develop new assays.
This platform – supplied as part of a partnership between Tecan and Waters to streamline mass spectrometry workflows for clinical laboratories – has all the hardware, labware and software scripts needed to perform SPE using 96-well Oasis® MAX µElution plates. Emma continued: “For most assays, patient samples and internal standards are pipetted into an initial 96-well collection plate on the workdeck of the Freedom EVO, then shaken for the mixing step. For scripts that include a protein crash step, we remove the plate and transfer it to a standalone Members of the endocrinology team (left to right): Somia Janjua, Anna Kowalka, Amal Bashir, Emma Walker, Sinead Reidy, Aileen Bulosan Collado and Anne Rogers
centrifuge. If the 96-well assay plates require conditioning prior to samples being added, the platform performs these steps while the collection plate is
Imperial College Healthcare NHS Trust is
spectrometry in 2008, when we
made up of five hospitals across north
converted from radioimmunoassay to
west London, UK, and treats in excess of
LC-MSMS for vitamin D testing. This was
1.5 million patients every year. The trust
originally to increase throughput, as well
also plays host to North West London
as to move away from the use of
Pathology, a consortium of laboratories
radioactive reagents. We initially
providing a wide range of pathology
continued to prepare vitamin D samples
services across its catchment area and
manually, using a liquid-liquid extraction
beyond. The endocrinology service,
approach, but the exponential increase
means that all we have to do is place a
based in the Clinical Biochemistry
in vitamin D test requesting we were
cover on the plate and transfer it from
Department at Charing Cross Hospital,
seeing at that time meant that it was not
the Freedom EVO to the mass
is an example of the specialty services
sustainable. This prompted our decision
spectrometer and begin the analysis.”
on offer, and provides advanced
to switch to a 96-well microplate format
diagnostic testing for a range of
and automated SPE (solid phase
“The Freedom EVO platform is intuitive
hormonal diseases.
extraction) for mass spec sample prep,
for the operator to use; its graphical
and we now use this approach for as
interface is compatible with a busy
LC-MSMS technology is at the forefront
many of our LC-MSMS assays as
clinical laboratory. The first assay to go
of the endocrinology laboratory’s
possible. In 2015, as part of a tender for
live using the Tecan system was our
workflow, as Dr Emma Walker,
a new mass spectrometry system, we
aldosterone testing, and we initially used
Consultant Clinical Scientist and Trust
acquired a Freedom EVO® workstation
a pre-programmed workflow. We often
Lead Healthcare Scientist, explained:
to help us further increase the efficiency
obtain basic ‘starter’ scripts from Waters
“We first began using mass
of our automated sample preparation.”
for new assays, then adapt these to our
24
TECAN JOURNAL 2/2017
being centrifuged. It then prompts the user to put the spun collection plate back onto the workdeck, and all the subsequent steps are carried out automatically, including sample addition, washing and elution into an LC-MSMS compatible solvent. This
CLINICAL BIOCHEMISTRY
MS sample prep workflow
Sample accessioning
Protein precipitation
Liquid-liquid extraction
Dilute and shoot Hydrolysis Positive pressure SPE or vacuum 96-well plate/ cartridge
Applications
Sample tracking
Chromatography and mass spectrometry
Extraction
Data integration LIMS
Drug screening and vitamins Therapeutic drug monitoring and drugs of abuse Steroids and catecholamine
Optional devices • Evaporator • Centrifuge • Sealer
specific requirements as we work
able to use to develop new assays and
through our own method validation in
widen our test repertoire. By
the laboratory. Several members of the
implementing an automated, integrated
laboratory team have been on training
approach, we have gone from a situation
courses to help us fully exploit the
where we didn’t have enough pairs of
potential of the system and we now also
hands in our laboratory, to one now
design and set up our own scripts. For
where we are using Tecan solutions to
example, we now offer a ‘core’ steroid
help us maximize what we can get out
panel, which includes testosterone,
of our mass spectrometry systems,”
androstenedione and
Emma concluded.
17-hydroxyprogesterone. We run this steroid panel assay two or three times a
All Tecan products mentioned are for research
week, with each full plate
use only. Not for use in clinical diagnostics.
accommodating around 80 patient samples – plus standards and controls – and it takes approximately 90 minutes to prepare for analysis.” “The walkaway automation offered by the Freedom EVO workstation has saved a lot of staff time in the laboratory compared to when we were doing manual liquid-liquid extraction. Using this platform to help streamline the flow of samples through our service has
U sing this platform to help streamline the flow of samples through our service has liberated staff time, which we have been able to use to develop new assays and widen our test repertoire.
To find out more on Tecan’s clinical laboratory solutions, visit www.tecan.com/diagnostics For more information on North West London Pathology’s laboratory services, including diagnostic endocrinology, go to pathology.imperial.nhs.uk
liberated staff time, which we have been
TECAN JOURNAL 2/2017
25
GENOMICS
Perfect synergy for molecular diagnostics Automation has an important
Molecular diagnostics is becoming
gastrointestinal diseases – and
role to play in molecular
increasingly common in laboratories
microfluidics and optics expert
worldwide, and this has driven a demand
Genewave in 2013 to form the present
to move from tedious, time-consuming
day organization. Juha Kirveskari, R&D
and labor-intensive manual processes to
Director at Mobidiag, explained: “Clinical
automated workflows. A further
laboratories performing stool diagnostics
throughput. Mobidiag has
consideration for clinical laboratories is
can be working with dangerous
developed a CE-marked,
the potential hazards associated with
pathogens. User safety is paramount,
automated platform for
handling dangerous pathogens, where
and molecular diagnostics can minimize
user safety is a priority.
the risk associated with these analyses.
diagnostic workflows, minimizing manual interventions to help enhance
nucleic acid extraction and PCR plate set-up, enabling high volume screening and antibiotic resistance testing for gastrointestinal pathogens.
However, when performed manually, the Mobidiag is a Finnish biotech enterprise
process is laborious and time consuming.
specializing in developing and marketing
We already had a semi-automated
novel, innovative solutions for molecular
platform, and wanted to streamline the
diagnostics of infectious diseases. The
process further, fully automating our
Espoo-based company was originally
suite of real-time PCR-based assays for
founded in 2000, merging with
gastrointestinal diseases to eliminate the
Amplidiag – a stool-based clinical assay
numerous manual procedures required
development company focused on
before and after extraction.”
The Amplidiag Easy R&D team (left to right): Mikko Vainio, Anniina Raitila, Juha Kirveskari, Sanna Laakso and Jaakko Kurkela
26
TECAN JOURNAL 2/2017
GENOMICS
T he automated system enables us to perform the extraction in two hours, from start to finish, producing two ready-to-run qPCR plates. Mobidiag worked with Tecan to develop
48 samples at a time, physically
On completion of the run, automated
the Amplidiag® Easy system for
separating them in a 96-well plate.
data analysis is performed, and the
automation of nucleic acid extraction
The automated system enables us to
results exported to LIMS. Manual
and PCR plate set-up for the Amplidiag
perform the extraction in two hours,
interventions are virtually eliminated.”
multiplex stool diagnostic tests for
from start to finish, producing two
gastrointestinal infections. “I knew from
ready-to-run qPCR plates. It is just not
“As a CE-marked instrument, the
clinical experience that the existing
practical to do this manually, as it is
configuration and coding of the system
technologies had limitations, and were not
quite a complex assay set-up. In theory,
are locked, but there’s still quite a lot of
ideal for our purposes. I’d used a Tecan
it would take a full day to perform, but it
flexibility for customization, such as
platform in an open configuration in a
would be almost impossible to maintain
tailored protocols for extraction or
clinical laboratory in the past, so I knew
the high level of concentration required
PCR set-up only. This offers additional
the technology and the software, and
to handle large numbers of samples
possibilities, for example, the
that the hardware was reliable and could
without making a mistake.”
implementation of in-house assay
withstand routine daily operation without
procedures. The system is now being
technical issues. I also knew that the local
“The first stage of the workflow is
distributed across Europe and other
service personnel are highly professional.”
transfer of the homogenized sample to
countries where CE marking is valid,
the laboratory in an automation-friendly
and we are expanding and developing
Juha continued: “We discussed our ideas
tube containing liquid medium. On
our service organization to ensure
with Tecan, and the company suggested
arrival, it is barcoded and the cap is
that laboratories have access to the
some very helpful features. The result
removed, then the entire extraction
technical advice and support they need.
is Amplidiag Easy, a versatile, fully
process is performed on the automated
It has been a very successful project,”
integrated platform operated through a
platform. The run is started via the GUI,
Juha concluded.
simple TouchTools™-based graphical user
which prompts the user to check that
interface (GUI). It includes a Te-MagS™
the set-up is correct, then the DNA
All Tecan products mentioned are for research
module for our magnetic bead-based
extraction and qPCR plate set-up are
use only. Not for use in clinical diagnostics.
assay technology, a heat block, and a
performed without the need for user
PosID™ barcode reader, which saves a lot
intervention. After manual heat sealing,
of user time by automatically scanning
the plate is moved to the CFX qPCR
all the sample tubes. The system can also
instrument (Bio-Rad) for processing,
To find out more about
accommodate all the consumables
while the sample data is transferred via
Tecan Partnering, visit
needed for our assays; reagent tubes,
either a USB stick or a LIMS. The CFX
www.tecan.com/partnering
extraction beads and PCR plates.”
system reads the plate barcode and imports all the sample data and
To learn more about Mobidiag,
“In a clinical laboratory, it is vital to avoid
protocols from the LIMS, starting the run
go to www.mobidiag.com
cross-contamination, and so we extract
automatically once the plate is in place.
TECAN JOURNAL 2/2017
27
IMMUNOLOGY
Luciferase comes to the devil’s rescue Wild Tasmanian devils are vulnerable to a facial cancer discovered in 1996 and identified as a transmissible tumor a decade later. The contagious disease originated in northeastern Tasmania and spread throughout the country, decimating the devil population and raising the real possibility of extinction. Scientists at the Menzies Institute for Medical Research, University of Tasmania, have pioneered research into the problem – drawing upon the latest developments in human immunology and bioluminescence cytotoxicity assays – in the hope of developing a vaccine to save the island’s iconic marsupial. The Tasmanian devil facial tumor
Andy Flies, a postdoctoral research
PD-L1 is upregulated in devil facial tumor
disease is responsible for as much as a
fellow at the University of Tasmania
cells in response to interferon-gamma –
90 percent decline in the population
supported by the Morris Animal
a cytokine released by activated T cells.
since 1996, proving fatal in nearly every
Foundation and a Commonwealth grant
The PD-L1 binds to PD-1 receptors on
case of infection. In 2014, a second
co-funded by Nexvet Biopharma,
the T cells, inhibiting the antitumor
genetically distinct and independent
explained: “An organism’s immune
immune response and enabling the
transmissible tumor was discovered with
system should launch an attack on any
tumor to evade detection. In short,
similar consequences – large lumps
foreign cells, which is what makes the
appear around the mouth and head that
transmissible tumors so intriguing. The
prevent eating and lead to starvation.
cancer cells are transferred as an
The recent discovery of some disease-
allograft when the devils bite one
“The big headlines in recent human
resistant devils suggests that the species
another, and our team is trying to figure
immunology have covered checkpoint
may be rapidly evolving, but there is still
out why there is no immune response. In
no devil immune response to the tumor
a recent publication,1 we showed that
in the majority of the population.
the key immune checkpoint molecule
the production of PD-L1 acts as an invisibility cloak.”
molecule inhibitors. We’re tracking what is working in human clinical trials, and trying to reverse-engineer and adapt it for our own use. Our latest publication compared key immune checkpoint molecules in nine different species, ranging from humans to bats to Tasmanian devils.2 Despite the last common ancestor of marsupial and placental animals occurring 162 million years ago, we found a remarkable level of similarity in key regions for these critical immune molecules, suggesting that some immunotherapy or vaccine approaches for humans might also work in Tasmanian devils. The lack of devilspecific antibodies has been a real obstacle to progress in this area. Using antibodies from other species has yielded only moderate success, so we have developed around 50 monoclonal antibodies of our own against seven or eight different targets. We’re starting to gather the necessary resources to carry out more advanced immunology, beginning at a similar point to human and mouse immunology 20 to 30 years
The research team at the Menzies Institute for Medical Research with actress and ambassador Bonnie Sveen (left) 28
TECAN JOURNAL 2/2017
ago with regard to the availability of species-specific reagents.”
IMMUNOLOGY
Andy continued: “To date, the chromium-51 release assay has been the gold-standard approach to studying cellular immune responses. Target tumor cells are labeled with the isotope and
I now use the Spark for all my ELISAs, due to its speed and the option to use different wavelengths.
incubated with effector cells, and the immune response can be determined by measuring radioactivity, as the cancer cells release the isotope when they are killed. Working with radioactive chemicals is a hassle, as there are many regulations and chromium-51 has a really short half-life of only 21 days; after a single month, half of a $500-dollar isotope stock has decayed and, if you encounter a delay, you end up wasting your money.” “There was a need and a desire to move on and find an alternative method, and that’s where the Spark® comes in. Our colleague, Nuri Guven, raised the funds to purchase the instrument, and I first became aware of the Spark’s great potential after I attended a Tecan Spark workshop organized at the Menzies Institute for Medical Research. Nuri’s previous experience with Tecan systems was very helpful in getting us up to speed on the instrument. In the new assay, I transfect the target tumor cells to express luciferase – an enzyme derived from the firefly – making them luminescent. The bioluminescence is proportional to the number of surviving tumor cells, so the cell survival rate can be measured by quantifying the luminescence. Although we have a few other readers in the department, they were not capable of carrying out this particular assay.” “The Spark software is pretty straightforward and the results from the first two tests confirmed that it was suitable for our needs. The cytotoxicity assay is really fast; running a whole plate takes about 90 seconds, and it is much better to run eight plates in 15 minutes than waiting for a chromium counter for two hours. I now use the Spark for all my
1) Flies Andrew, S et al. PD-L1 Is Not
ELISAs, due to its speed and the option to
Constitutively Expressed on Tasmanian Devil
use different wavelengths. In the future,
Facial Tumor Cells but Is Strongly Upregulated
we plan to purchase the Humidity
in Response to IFN-γ and Can Be Expressed in
Cassette and Te-Cool®, which will enable
the Tumor Microenvironment. Frontiers in
us to incubate cells in the reader, allowing
To find out more about the Spark’s luminescence optics, visit www.tecan.com/spark
Immunology, 2016, 7, 513.
To learn more about Andy Flies’
us to run kinetic assays with automatic
2) Flies Andrew, S et al. Comparative Analysis
injection overnight. Our ultimate hope is
of Immune Checkpoint Molecules and Their
research, visit
that these new assays will help us to
Potential Role in the Transmissible Tasmanian
utas.edu.au/profiles/staff/
develop a vaccine but, whatever the
Devil Facial Tumor Disease. Frontiers in
menzies/andrew-flies
outcome, our research has implications
Immunology, 2017, 8, 581.
for our understanding of human and
All Tecan products mentioned are for research
veterinary cancer,” Andy concluded.
use only. Not for use in clinical diagnostics.
TECAN JOURNAL 2/2017
29
DRUG DEVELOPMENT
Expanding the experimental space Biomanufacturing requires careful separation of the molecule of interest from other cellular products to ensure the quality and stability of the final product. This is even more important for the manufacture of biotherapeutics, as the presence of unwanted molecules in pharmaceutical preparations can affect the efficacy and safety of biologically-derived drugs. FUJIFILM Diosynth Biotechnologies is using automated chromatography condition screening to generate more data in less time, helping to improve the performance of purification processes and accelerate biological drug manufacturing.
Biotherapeutics are now the key growth
derived medicines, and works with some
effects. This process is complex even
area for pharmaceutical manufacturers
of the most prominent biotech
for large macromolecules derived from
globally, but the complexity of
developers and pharmaceutical
mammalian cell lines, such as
manufacturing biological drugs means
companies in the industry.
monoclonal antibodies (mAbs), but is
that there are significant costs
even more difficult when using
associated with the development of
The company’s UK site near Billingham
microbial expression systems. Column
scalable production and purification
specializes in microbial expression
chromatography is the method of
processes. Rather than cultivating
platforms – mainly yeast and E. coli –
choice for most biomanufacturing
in-house expertise in production
and has developed a high throughput
applications, and so we need the ability
bioengineering and purification, many
chromatography workflow to help
to screen a wide range of
pharma and biotech companies now
select and optimize its purification
chromatography resins and parameters
partner with contract development and
processes. Jonathan Rapley, Staff
to ensure high purities and yields for
manufacturing organizations (CDMOs)
Scientist in High Throughput Process
our pharma customers. This is both
for the production of new
Development, explained: “The cGMP
labor intensive and time consuming to
biotherapeutics. FUJIFILM Diosynth
manufacture of biotherapeutics requires
perform manually and so, historically,
Biotechnologies has over 20 years of
the careful separation of the molecule
the scope of these screens has been
experience in process development and
of interest from other cellular products
limited by the development timelines
cGMP manufacturing for biologically-
and components to avoid off-target
of each project.”
Automated chromatography condition screening helps to generate more data in less time
30
TECAN JOURNAL 2/2017
DRUG DEVELOPMENT
The Freedom EVO 200 offers complete automation of resin and condition screening
T he Freedom EVO platform is a truly open laboratory automation solution; you can make it do almost anything.
Using design of experiment principles,
almost anything if you put the time into
we can fully explore the experimental
programming it.”
window, instead of having to investigate a far smaller number of combinations
“Both myself and a colleague have been
of resins and buffer conditions based
on Tecan training courses to further
on previous experience. This ensures
explore the potential of these
more complete data sets and helps
workstations, and we are now the ‘super
our project managers to make better
users’ who program new applications
informed decisions regarding production
and support other users, which is
processes, ultimately benefitting
working really well. As more and more
our customers.”
people have begun to understand the
“Automation of chromatography
capabilities and advantages of the
condition screening is an obvious way
“The increase in the number of
systems, demand is really ramping up;
of increasing the number of parameters
experiments we can perform has
we’re increasingly running them
that can be investigated with the
obviously led to a significant rise in
overnight and at weekends to meet
available time and resources, and so in
downstream analyses, as each screen
demand,” Jonathan concluded.
2015 we looked at the options available
generates six 96-well plates of samples.
for this application. The combination of
To help us deal with this, we have also
All Tecan products mentioned are for research
the Freedom EVO® liquid handling
invested in a Freedom EVO 100
use only. Not for use in clinical diagnostics.
workstation and RoboColumns®
platform. The system is configured for
(Repligen) was an ideal solution for our
ELISA testing, but the open architecture
needs, as this would allow us to run eight
of the Freedom EVO workstation means
pre-packed miniature chromatography
that, as we have become more familiar
columns in parallel, without the need for
with the software, we have also begun
any manual intervention. We invested in
using it to automate sample preparation
a Freedom EVO 200 platform, giving us
for other analyses, such as HPLC or
enough worktable space for all the
capillary electrophoresis. This broad
reagents required to screen eight
range of capabilities is a real benefit of
different resins at three different pHs per
Tecan systems. Unlike many other
run. The increase in capacity offered by
instruments, which are restricted to
For more information on FUJIFILM
this set-up means that we are now able
single applications, the Freedom EVO
Diosynth Biotechnologies, go to
to screen far more parameters for each
platform is a truly open laboratory
www.fujifilmdiosynth.com
project within the same timeframe.
automation solution; you can make it do
To learn more about Tecan’s chromatography solutions, visit link not working. Use this one instead: www.tecan.com/ proteinpurification
TECAN JOURNAL 2/2017
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 +31 183 44 81 74 Singapore +65 644 41 886 Spain +34 935 95 2531 Sweden +46 8 750 39 40 Switzerland +41 44 922 81 11 UK +44 118 9300 300 USA +1 919 361 5200 Other countries +43 62 46 89 33 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 To register for the Tecan Journal please go to www.tecan.com/journal © 2017 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