TJ Edition 2/2015
Tecan Journal
Life Sciences and Diagnostics
Automated PBMC isolation for biobanking pages 10 - 11
Streamlining medical genetics with the aid of automation pages 12 - 13
Stressing the importance of quality assays pages 22 - 23 28
Genomics solutions for marine biology pages 16 - 17
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Welcome to the summer issue of the Tecan Journal
The first few months of 2015 have been an exciting time for Tecan, with very positive feedback about our Fluent™ laboratory automation solutions and the new Spark™ 10M multimode reader coming from across the market. We are particularly proud of how the Fluent platform has been received by early users, with some customers already coming back to order additional systems. In this issue, we take a look behind the scenes at what has made this instrument such a success, with members of the Fluent team offering an insight into how pioneering concepts translate into application-oriented solutions. This flow of innovative new products is set to continue, with the launch of new models and new features for both platforms throughout the year. For the clinical diagnostics community, the 2015 AACC Annual Meeting in Atlanta this July will provide the perfect opportunity to learn how we can support your automation needs. Visitors to Tecan on booths #3636 and #3652 will have the opportunity to explore two of the current hot topics in diagnostics: the increasing role of automated sample preparation in genomics, and the latest developments in the field of immunodiagnostics. We will be demonstrating some exciting automated solutions for both these applications, with our colleagues from IBL International showcasing how our portfolio of immunodiagnostic assay kits can be efficiently automated on liquid handling workstations. You can learn more about the benefits of combining these high quality assays with our powerful automation solutions in this Tecan Journal, with a story from a long-term customer who is using this approach to provide robust and reliable specialist testing for psychology research. Elsewhere in the issue, we take a look at the latest advances in disease-oriented biobanking at IBBL in Luxembourg, and how Spanish energy producer Repsol is using automation to accelerate research into alternative energy sources. I hope you enjoy this issue, David Martyr, CEO
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CEO WELCOME TECAN JOURNAL 2/2015
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page 5
A complete solution for automated solid phase extraction 2
CEO Welcome
4 Spark 10M already lighting up research
pages 8 - 9
5 Total automation of SPE for toxicology and biochemistry applications 5 Application guide for Tecan plate washers offers an easy route to optimized results 6 - 7
Fluent – behind the scenes
8 - 9
Perfect chemistry
10 - 11 Automated PBMC isolation for biobanking 12 - 13 Streamlining medical genetics with the aid of automation 14 - 15 Back-to-back success for bioanalytics 16 - 17 Genomics solutions for marine biology pages 18 - 19
18 - 19 Driving oncology drug development forward 20 - 21
Developing the fuels of the future
22 - 23 Stressing the importance of quality assays 24 - 25 A simple(x) approach to bioprocessing 26 - 27 Freedom to evolve 27
Leading the debate
28 Events pages 24 - 25
Exploring novel bioprocessing applications
CONTENTS TECAN JOURNAL 2/2015
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Spark™ 10M already lighting up research Since its launch at SLAS 2015 in February, the Spark 10M has gained interest from across the life sciences community, with researchers around the world beginning to explore the advanced capabilities of this exciting new system for applications including DNA quantification, microbiology research and cell-based assays. To further extend the flexibility and appeal of this powerful system, Tecan has introduced a number of new features, including a cuvette port for absorbance measurements and an enhanced detection module offering straightforward performance of immunoassays using Alpha Technology. The enhanced detection module makes it easier than ever before for customers to perform AlphaScreen®, AlphaLISA® and AlphaPlex™ protocols. This option combines a high power laser light source with both low-pass and high-pass filters, using pre-optimized filter settings to ensure exceptional assay performance in 96- and 384-well microplate formats. The introduction of a cuvette port further
increases format flexibility, allowing
this feature helps to avoid loss of data by
ultra-fast absorbance measurements
automatically adjusting the gain settings if
using Tecan’s patent-pending High-Speed
signal saturation is reached. The software
Monochromators.
then automatically correlates the data across different gain settings, allowing users
The instrument’s SparkControl™ software
to view the entire dataset in a single graph.
has also been enhanced, with a number
In addition, the introduction of unique,
of new and improved features to
one-click applications – including
streamline routine activities and maximize
absorbance reading, cell counting, cell
productivity. An extended dynamic range
viability, nucleic acid quantification and
option enables both low and high gain
nucleic acid labeling efficiency – makes it
measurements within a single microplate,
easy to perform optimized measurements,
allowing complete data sets to be obtained
offering rapid, consistent results at the
without compromising on sensitivity for
touch of a button.
low signal strength measurements. This is complemented by an automated gain
Dr Tobias Kiesslich from the Institute
regulation function for fluorescence-based
of Physiology at the Paracelsus Medical
measurements. Ideally suited to kinetic
University in Salzburg, Austria, is a long-
assays and long-term cell-based studies,
term Tecan collaborator, and has already gained practical experience with this groundbreaking multimode microplate reader within his laboratory’s daily research: "The Spark 10M is straightforward to use and program, offering a great range of applications and measurement modes. The innovative cell counting module is particularly useful, allowing the instrument to make the step up from a microplate reader towards full automated imaging. The SparkControl software provides a full range of functions which are easy to use, with a clear modular programming concept to make designing and running experiments very simple. Together with features such as gas and temperature control, automated lid handling, dual injectors and evaporation protection, the Spark 10M offers flexible and versatile experimental design, even for cellbased applications." To find out more about the Tecan’s Spark 10M multimode reader, visit www.tecan.com/ignite
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PRODUCT NEWS TECAN JOURNAL 2/2015
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Total automation of SPE for toxicology and biochemistry applications Tecan has launched the Freedom EVO® positive pressure SPE workstation to provide a complete solution for automated solid phase extraction in clinical research, toxicology, forensic and hospital laboratories. SPE is a common sample preparation technique in many laboratories, but current workflows are largely based on manual processing. This can cause major throughput and quality control challenges, as the higher error rates associated with manual protocols can lead to repeat testing and increased costs. The Freedom EVO positive pressure SPE workstation enables labs to reproducibly automate extraction of a variety of clinically important analytes – including drugs of abuse, vitamin D and steroids – offering reproducible extraction, reduced error rates, lower solvent consumption and significant time savings, with full data traceability to ensure regulatory compliance. The workstation can extract up to eight samples in parallel using most industry
standard 1, 3 or 6 ml SPE cartridges, with individual control of pressures and flow rates in each channel to ensure superior analyte recovery compared to vacuum-based techniques. Fully automated barcode reading enables hands-free implementation of LIMSgenerated worklists, ensuring a high level of sample traceability and virtually eliminating the risk of errors. The system is controlled via an applicationspecific TouchTools™ graphical user interface, offering simple ‘click and run’ protocols with minimal user training requirements. This ease of use, combined with reliable and costeffective extraction, offers a unique, end-toend SPE solution that is already benefitting laboratories around the world1.
To find out about Tecan’s mass spectrometry sample preparation solutions, visit www.tecan.com/lcms 1 Fernández et al. J Anal Toxicol, 2014, 38(5), 280-288.
The system can be used with most 1, 3 or 6 ml SPE cartridges
Application guide for Tecan plate washers offers an easy route to optimized results Tecan’s recently published Application guide for Tecan plate washers puts information at your fingertips, allowing users of the popular HydroSpeed™ and HydroFlex™ instruments to optimize their assays more easily. Whether you are performing ELISAs, vacuum filtration, magnetic bead- or cell-based assays, the guide contains everything you need to know about Tecan washers. The guide includes an overview of the features and highlights of each instrument – for example extra gentle cell washing with the HydroSpeed’s Cell Protection™ function – enabling users to select the most appropriate system for their particular application, as well as references to supporting literature and application notes. Instant access to all the knowledge required to precisely control critical wash parameters – such as vacuum
power, aspiration position, and dispensing rates and positions – helps users to improve assay consistency by avoiding detachment of cells, beads and antibodies during wash steps. The Application guide for Tecan plate washers contains such a wealth of essential
information that it is sure to prove invaluable in the laboratory. For more information, or to download a copy of the guide, visit www.tecan.com/cell-protection
The new application guide offers useful hints and tips to improve washing performance
PRODUCT NEWS TECAN JOURNAL 2/2015
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Fluent™ – behind the scenes The launch of the latest Fluent laboratory automation solution has brought the advantages of this application-oriented platform to a host of new users, offering walkaway compound management and assay plate generation to complement the existing end-to-end automation of cell-based and biochemical assay protocols. To learn more about the benefits of the Fluent workstation’s innovative liquid handling automation concept, we spoke to some of the team responsible for its development.
At the heart of the Fluent platform is a unique instrumentation concept, as Kevin Moore, manager of the liquid handling products and applications team, explained: “Resource pressures mean that many laboratories no longer have the time to perform application development in house. As a result, they are increasingly looking for ‘ready to run’ solutions that have been developed and validated at the vendor level. From the outset, the design brief for the Fluent project was to create a liquid handling workstation that focused on our customers’ applications, building on the success of our market-leading Freedom EVO® series of open architecture workstations.” “Ensuring that these integrated solutions provide straightforward set-up and walkaway performance, while still offering the flexibility to fit individual laboratory workflows, was the real challenge. By working with expert academic and industry partners, we have been able to guarantee that each Fluent laboratory automation solution offers superior precision, throughput and walkaway time for everyday activities.” Bronwen Forster, senior product manager for liquid handling, took up the story: “This was certainly an exciting project. It covered so many different aspects of the laboratory workflow, from the most commonly used sample formats through to the final goals of a laboratory’s research. From a hardware perspective, one of our main aims was to increase modularity and flexibility, making it easier to build a system that meets a customer’s specific requirements – and can adapt as those needs change – without
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PRODUCT NEWS TECAN JOURNAL 2/2015
creating bespoke solutions. The instrument’s Dynamic Deck™ is an example of this, providing greater deck capacity and unique multidimensional access for the integration of virtually any combination of modules and devices on, in, around or below the workdeck.” “As we were creating an entirely new platform, we also had the opportunity to develop new electronics and motor control hardware to ensure straightforward, robust integration of both Tecan and third-party devices. This not only enhances speed and control for individual operations, it also provides greater stability and enables introduction of ‘teach-free’ hardware modules and Zero-G one-touch teaching for the set-up of external devices. Ease of maintenance is almost as important as instrument set-up and operation, and the Fluent platform is
designed to allow all maintenance tasks to be performed from the front of the system. This may seem like a small thing but, with an increasing trend towards liquid handling platforms being fully enclosed for applications such as cell-based assays, this can make servicing activities much faster, minimizing downtime for busy laboratories.” Hal Wehrenberg, who led the team developing the new FluentControl™ software, added: “We’ve been doing laboratory automation for a long time, and we’re already a market leader in terms of usability and functionality, so it was a lot of fun as a company to ask ourselves ‘What can we do even better?’ This was very much the design principle for FluentControl, and probably the biggest innovation was to acknowledge that what the day-to-day operator needs is different from what the system programmer
A multidisciplinary team developed the Fluent workstation hardware…
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“…it was a lot of fun as a company to ask ourselves ‘What can we do even better?’”
wants. We chose to focus on both of these, targeting them directly with separate solutions so that we didn’t have to compromise for either.” “We started by creating a dedicated user interface that’s optimized for the daily laboratory workflow. Taking design cues from smart phones and tablets, it is very intuitive and walks the user through maintenance or assay set-up, making it as easy as possible for operators to run experiments. Unlike the standard touchscreen interfaces now available across the industry – which use the same commands but ‘bigger buttons’ – we have
special commands for embedding pictures and text, which we believe is completely unique in the marketplace. This allows laboratories to take their SOPs – which might be in a binder that you hope people read – and integrate just the parts needed for the run the user is about to perform into the instrument set-up process. It’s right there for every single user and application, and you can fully customize it to your needs, adding photographs of the workdeck or even videos of, for example, how to remove a plate seal correctly.” “From the other perspective, we didn’t want to ignore the needs of advanced users and
programmers, and so we have a separate programming interface. Continuing the theme of ease of use, one of the most exciting new concepts for this interface is the introduction of Smart Commands. These are simple programming tools to take care of those activities that are part of every protocol, such as reagent distribution or sample transfer. Instead of writing out a script line by line – sometimes up to 30 lines of code – we’ve created single line commands that allow you to just add the parameters you need. We still have the advanced functionality to cover virtually any task you can think of, but we’ve made it faster to write basic protocols. We’ve also worked very closely with both our application specialists and beta testers to make sure that these Smart Commands are still useful for expert users, not just basic level programmers. It was an interesting design challenge, but I’m really proud of how we’ve pulled it off; maintaining the robustness, flexibility and power that people expect from a Tecan liquid handling platform, while making it easier than ever before to program and use.” To find out more about Tecan’s Fluent laboratory automations solutions, visit www.tecan.com/fluent
…working closely with the FluentControl software team
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Perfect chemistry Italy’s Reparto Investigazioni Scientifiche has invested in an HID EVOlution™ to aid analysis of DNA from crime scene samples, providing the process security that is essential in any forensic science laboratory.
“I can always be sure that I am reading all the documentation relating to a particular sample.”
The RIS team with the HID EVOlution
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FORENSICS TECAN JOURNAL 2/2015
Reparto Investigazioni Scientifiche (RIS) in Rome is one of four scientific investigation departments of the Italian military police, the Carabinieri. The laboratory comprises 20 technicians and six supervisors, and is responsible for the forensic analysis of DNA evidence from crime scenes across central and parts of southern Italy, as section commander of biology Andrea Berti explained: “We analyze a wide range of sample types that can potentially be found at a crime scene – such as blood, saliva and contact traces – for DNA. We also carry out research projects, helping to develop and test new products for DNA and RNA analysis. As part of this research, we have worked with the European DNA Profiling (EDNAP) group analyzing RNA samples, the University of Rome ‘Foro Italico’ to test forensic applications of next generation sequencing, and the University of Florence for anthropological analysis of mitochondrial DNA.” “Typically, we handle around 100 cases – in the region of 400 to 500 samples – in a single month, a large number of them low copy number DNA samples. Like many laboratories, we need as much walkaway time as possible,
and believe that any manual operation that can be performed just as well – or better – using automation should be transferred to an automated platform. Previously, we relied on a small automated platform for sample extraction, but this had a very limited capacity, and so we invested heavily in higher throughput systems for our DNA analyses, installing the HID EVOlution alongside instruments from other manufacturers to give us the flexibility to run automated protocols for the entire range of chemistries used in forensics.” Andrea continued: “The HID EVOlution was chosen because it is a system that I know well, and which is extremely robust. All the features and applications of the instrument were explained to us by the Tecan team, and the collaboration between Tecan and Life Technologies was a great advantage, meaning that validated, optimized protocols were available for the kits we use: the PrepFiler® Automated Forensic DNA Extraction Kit, the Quantifiler® Duo DNA Quantification Kit, and the AmpFℓSTR® NGM™ Kit. The system was installed during summer 2014, and specialists from Tecan and Life Technologies helped
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us to set up the platform. This support was really important, ensuring that all our files are correctly imported and exported for real-time PCR quantification and DNA normalization. Training in the use of the instrument was also provided; this was very professionally delivered, with the Tecan technician working hard to solve any difficulties quickly. We now have two members of staff who are specialists in the use of the system and understand the platform perfectly, enabling them to help their colleagues carry out DNA analyses.” “In any laboratory, and especially accredited laboratories, process security is essential. Automation allows us to process a lot of samples, and all the relevant information – including sample volumes, extraction protocols and quantification methods – must be recorded. The HID EVOlution offers complete sample tracking, helping to increase the quality of the information and documentation generated for each sample, which is so important. It allows the laboratory to follow the progress of a sample through the entire workflow, store the data, and print a report, making data management so much easier; I can always be sure that I am reading all the documentation relating to a particular sample.” “RIS is an ISO 17025 accredited laboratory and, as such, must be very clear about the technologies and methods it uses for DNA analysis. Since the HID EVOlution was installed, we have validated the system in our laboratory, analyzing different sample types to demonstrate the performance of the instrument. The results were compared to our other, already accredited, methods and the data analyzed, and we are really pleased with the results. The validation report was accepted by the accreditation committee and, since December 2014, the Tecan workflow has been included in our schedule of ISO 17025 accredited methods,” concluded Andrea. To find out more about Tecan’s HID EVOlution, visit www.tecan.com/hid To find out more about RIS, visit www.carabinieri.it/arma/oggi/indaginiscientifiche
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Automated PBMC isolation for biobanking Since its inception in 2008, IBBL has been a biobanking pioneer, developing a high quality repository of samples to support biomedical research into a variety of diseases. Using a Freedom EVO® workstation, the center has developed an automated workflow for the processing and cryopreservation of a wide range of biological materials, and has recently added viable peripheral blood mononuclear cells to its standard whole blood extraction portfolio.
IBBL (Integrated BioBank of Luxembourg) is at the forefront of disease-oriented biobanking, using cutting-edge automation technologies to build up a large collection of disease-specific samples for research into new approaches for the diagnosis and treatment of a wide range of conditions. Working closely with the neighboring Centre Hospitalier de Luxembourg, IBBL catalogs and cryogenically preserves biological materials – including tumor samples, serum, blood, plasma, urine and feces – from individuals with specific diseases, for national and international projects. IBBL has a strong history of working in partnership with Tecan, and has already developed a number of innovative automated sample preparation protocols using a Freedom EVO 200 platform (Tecan Journal, Issue 3, 2012). The center’s latest project – to automate extraction of viable peripheral blood mononuclear cells (PBMCs) from whole blood – further demonstrates the advantages of IBBL’s automated approach. Dr Fay Betsou, Chief of Biospecimen Science, explained: “PBMCs – primarily lymphocytes and monocytes – are of increasing interest for vaccine development and validation, and we have seen a steady increase in requests from our collaborators within the pharmaceutical industry for PBMC-based functional assays. These blood cells are traditionally extracted manually using a Ficoll-Paque method. This gradientbased technique is both laborious and time consuming, taking a technician
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BIOPROCESSING TECAN JOURNAL 2/2015
Left to right: Gaël Hamot, Conny Mathay and Fay Betsou with the Freedom EVO workstation
approximately four hours to isolate PBMCs from 10 samples.” IBBL has developed a straightforward automated protocol for PBMC extraction based on the use of standardized citrate anticoagulant tubes (BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate). By ensuring the same fill volume in each tube, the team has been able to identify the optimum pipetting height following centrifugation, allowing efficient isolation of PBMCs while minimizing the number of granulocytes present in the final preparation. Fay continued: “Unlike the buffy coat extraction process, which
uses the Freedom EVO’s Tube Inspection Unit to identify the optimal pipetting height, this protocol is based on the mean hematocrit of the general population. Despite this, the recovery rates are comparable to the manual process – 61 % recovery with the automated process compared to 69 % when performed manually. This is very satisfactory for our needs and, more importantly, cell viability is virtually unaffected – 74 % compared to 79 % for manual processing.” Centrifuged, decapped CPT tubes are placed on the Freedom EVO workstation, and the plasma-suspended PBMCs are transferred into intermediate tubes, which are respun using
TJ Schematic input/output of the blood workflow 10 ml EDTA collection tube centrifuged and recapped
Script A
8 ml CPT tube centrifuged and recapped
Mixing tube to ensure homogeneity of aliquots
Mixing tube to ensure homogeneity of aliquots
Script B
Based on study aim
Mixing tube to ensure homogeneity of aliquots
10 ml CAT collection tube centrifuged and recapped
Mixing tube to ensure homogeneity of aliquots
Spin in embedded centrifuge 12 x 220 µl plasma 0.5 ml matrix tube
12 x 220 µl plasma 0.5 ml matrix tube
12 x 220 µl plasma 0.5 ml matrix tube
950 µl buffy coat + 3 ml water + 3 ml lysis buffer 15 ml conical tube
850 µl buffy coat + 850 µl lysis buffer FluidX 2 ml cryovial
PBMC in 10 ml PBS 15 ml conical tube
12 x 220 µl plasma 0.5 ml matrix tube
1.6 ml PBMC suspended in Cryostore10 FluidX 2 ml cryovial 100 µl buffy coat + 700 µl lysis buffer 96-well, deep well plate
IBBL’s blood sample processing workflow
“Recovery rates are comparable to the manual process… and, more importantly, cell viability is virtually unaffected.” the platform’s integrated centrifuge. The supernatant is subsequently discarded and the PBMC pellet is resuspended in cryopreservation medium in cryotubes. The processed samples are placed into cryostorage, first in a progressive rate freezer to avoid damaging the cells, and then into liquid nitrogen for long-term storage. “Best practice for cryopreservation is to normalize the cell concentration in each vial, however this is not possible using our standard workflow as cryopreservation medium is toxic at room temperature, and so samples need to be frozen quickly to maintain cell viability. This is not generally an issue as the automated process is very reliable and reproducible, however the Freedom EVO gives us the flexibility to resuspend cells in wash buffer
instead of cryopreservation medium if precise cell concentrations are required for specific projects.” The PBMC protocol has now been fully validated1 and implemented as part of IBBL’s standard sample collection workflow, allowing it to be run alongside a number of other scripts, such as the extraction of the buffy coat from blood. “Having implemented this protocol as part of a composite script, we can now automatically process different specimen types in parallel. Although high throughput is not our primary focus, the Freedom EVO offers walkaway processing of 24 patient samples in under three hours, allowing the technician to perform other activities while it is running,” Fay concluded. 1 Hamot et al. Method Validation for Automated Isolation of Viable Peripheral Blood Mononuclear Cells. Biopres Biobank, 2015, in press.
To find out more about Tecan’s biobanking solutions, visit www.tecan.com/biobanking
The PBMCs are extracted from centrifuged whole blood
To learn more about IBBL, go to www.ibbl.lu
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Streamlining medical genetics with the aid of automation With an ever-increasing workload, Germany’s Munich Leukemia Laboratory has automated its cytogenetics protocols on five Freedom EVO® workstations, enabling leaner, standardized, more efficient processing and minimizing the potential for human error.
The Munich Leukemia Laboratory
“Historically, extensive diagnostic testing
(MLL) provides a comprehensive range
was not necessarily performed, simply
of diagnostic tests to hospitals and
because there were no available treatment
doctors across the country, offering
options. However, medical knowledge has
cytogenetics and fluorescence in situ
advanced and treatments have improved,
hybridization (FISH), molecular genetics,
and now increased testing makes perfect
cytomorphology and immunophenotyping
sense. Usually, different diagnostic tests
services. The laboratory continually
are performed at separate laboratories, and
refines and streamlines its procedures
the results are collated and interpreted by
to cope with growing sample numbers,
a patient’s doctor. In contrast, each of MLL’s
and automation plays a key role in this,
four co-founders has a different background
creating standardized, efficient procedures
and specialty, and so we have developed a
and minimizing the potential for human
complete diagnostic and monitoring service
error. Head of cytogenetics and FISH, Prof
offering a wide range of tests under one
Dr med Claudia Haferlach explained:
roof. Currently, we handle more than 60,000
MLL uses its Freedom EVO platforms to improve efficiency and save time compared to manual processes
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CLINICAL DIAGNOSTICS TECAN JOURNAL 2/2015
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samples a year but, with the number of test
far less risk of processing errors. It also has
requests continually increasing, we have
the advantage of relieving staff of tedious,
turned to automation to streamline our
repetitive manual work, leaving them free to
workflows.”
carry out other more stimulating tasks such as research and the implementation of new
MLL has established automated Sanger and
technologies. Interestingly, when technicians
next generation sequencing protocols for
join the laboratory straight from school, they
molecular analysis, performing semi-
want to work manually because they think
automated isolation of mononuclear cells
it is boring to just push buttons. However,
by Ficoll separation on a Freedom EVO
after doing lots of manual pipetting, they
workstation, followed by RNA and DNA
soon change their mind and appreciate
preparation on a Roche MagNA Pure™ 96
sophisticated automation! Cytogenetics
System and PCR set-up on further
also involves a great deal of cell cultivation,
Freedom EVOs. In addition, automation of
routinely requiring sample preparation over a
the key stages of the laboratory’s tumor-
weekend. This can now be better automated
based cytogenetics and FISH workflows has
as well, which staff really appreciate.”
been implemented on its Freedom EVO workstations, tailoring each individual system
“Quality is crucial, and whenever we invest
to a specific stage of the process, such as
in new technology we evaluate a number of
hypotonic and fixation treatment of cell
companies working in that field, choosing
cultures for chromosome banding analysis, or
the one that performs the best. We selected
the pipetting of fixed cell suspension drops or
Tecan for the performance of its instruments
various FISH probes onto slides. Each platform
and its excellent quality assurance system
is equipped with a Liquid Handling Arm
and, after discussing the best way to transfer
and/or a Robotic Manipulator Arm, as well as
our existing manual processes onto the
any additional integrated modules required
Freedom EVO, worked with the company to
for its particular step of the workflow. After
automate our procedures. Having a good
slide generation on the Freedom EVO systems
working relationship with your supplier
and hybridization, FISH analysis is carried
is invaluable, as this is a two-way process
out using fully automated MetaCyte
requiring input from both sides; the
(MetaSystems) microscopy systems, with
laboratory needs to explain exactly what
electronic sample tracking via barcodes.
it would like the system to do, while the
“We are really satisfied with the Freedom EVO systems, which help us meet turnaround times and provide a high quality, costeffective diagnostic service.” when you work with skilled people,” concluded Prof Haferlach. To find out more about Tecan’s clinical diagnostics solutions, visit www.tecan.com/ clinicaldiagnostics To learn more about MLL, visit www.mll.com
supplier has an in-depth knowledge of the Prof Haferlach continued: “Some years ago,
instrument’s capabilities and knows how
all our cytogenetics and FISH workflows were
best to implement the necessary procedures
performed manually. We wanted to automate
on the workstations. Tecan listened to us,
these processes, improving efficiency, saving
took our comments on board, and used its
time for our technicians and allowing
knowledge to tailor the Freedom EVO systems
unattended overnight processing. No matter
to our workflow. We are really satisfied with
how good your workflows and staff are,
the systems, which help us meet turnaround
mistakes happen. Automation will always be
times and provide a high quality, cost-effective
more reproducible, generating better quality
diagnostic service. Collaborating with Tecan
results than manual procedures; there is
was good; you certainly notice the difference
Straightforward set-up and walkaway operation are key benefits of the Freedom EVO workstations
CLINICAL DIAGNOSTICS TECAN JOURNAL 2/2015
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Back-to-back success for bioanalytics Novartis’ Drug Metabolism and Pharmacokinetics (DMPK) Biologics Division, based in Basel, Switzerland, has developed a flexible, fully automated workflow for blood serum sample preparation and ELISA processing by combining two Freedom EVO® 150 workstations. The back-to-back configuration provides the Bioanalytical Group with the features and throughput to support a wide range of automated ELISAs, as well as offering semi-automated sample preparation for other analytical techniques.
Left to right: Lionel Florsch, Sophie Lieb, Christophe Zickler and Ulf Klein with one of the group’s Freedom EVO systems
Biologics – therapeutic preparations consisting of antibodies, proteins or other biologically-derived macromolecules – are of ever-increasing importance in the pharmaceutical industry, but their effects on the body are often more subtle or complex than traditional small molecule drug compounds. The Bioanalytical Group of the Novartis DMPK Biologics Division in Basel was
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DRUG DISCOVERY TECAN JOURNAL 2/2015
set up specifically to identify these effects in preclinical and clinical samples, measuring pharmacokinetics, pharmacodynamics and, crucially, immunogenicity. Dr Ulf Klein, Laboratory Head in the Bioanalytical Group, explained: “Across the industry, biologics portfolios are expanding rapidly compared to small molecule pipelines, and there are evermore sophisticated drug modalities
coming in. Where biologics used to be almost exclusively monoclonal antibodies, there is now a broad spectrum of antibody fragments, therapeutic proteins, peptides and other formats in development for various indications.” “This increasing complexity, combined with rising sample numbers, meant that
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“The Tecan instruments are almost like extra analysts.”
we needed to look for more efficient ways of performing routine analyses, giving us more time for the development of more sophisticated assays. Automation was one of the options, and so we looked at the various systems on the market. Tecan’s Freedom EVO seemed the best fit for our needs, with intuitive software that would enable us to develop and modify assay protocols as required, while still meeting our need for a validated system to operate in a GLP-/GCPregulated environment.” Ulf continued: “The Tecan team worked closely with us to understand our existing workflow and needs in terms of flexibility and integration of third-party devices. Once we had established the basic specification, Tecan developed the system around two back-to-back Freedom EVO 150 platforms, one to perform sample identification, distribution and dilution, and a second to run ELISAs. This interconnected set-up provides maximum process security while still allowing efficient use of the available deck space.” “Most of our ELISA workflows are fully automated; the operator simply loads the decapped sample tubes onto the system and starts the process. Each sample tube has a barcode which is scanned automatically, and then the system compares the barcode to
the assay input list derived from our LIMS. Once matched, samples are distributed into 96-well plates and diluted according to information in the LIMS. Tecan’s graphical user interface is very powerful, making it easy to perform these tasks while offering exceptional flexibility, for example we can select a dilution range from 1:1 to 1:1.5 million. Following reformatting and dilution, we either take the 96-well plates off the system and continue the analysis offline – our semi-automated workflow – or the plates are automatically transported through to the second Freedom EVO instrument to run the ELISAs.” “Depending on the project we’re supporting, the ELISA protocol can involve simply incubating the sample with an antibody, then washing and reading with the integrated third-party washer and multimode reader respectively, or several wash and incubation steps using multiple reagents. The incubation temperature and times can also vary significantly between assays, which can be easily adjusted. The Freedom EVOware® scheduling software also efficiently arranges the various ELISA steps, allowing us to run several plates without any loss of performance.”
and technical support was invaluable in achieving this. We can now support a number of ELISA protocols, as well as preparing samples for analysis by other methods, such as mass spectrometry.” “Overall, we have been very pleased with the liquid handling performance and reliability of the Tecan solution, so much so that we have now purchased a second system to allow us to automate more of our sample processing and ELISAs. The Tecan instruments are almost like extra analysts – ones that don’t mind doing repetitive tasks or working overnight – so we develop our assays manually, then ‘hand them over’ to the Tecans for routine testing. This ensures reproducible assay performance and a complete audit trail, as well as giving us more time for assay development,” Ulf concluded. To find out more about Tecan’s drug discovery solutions, visit www.tecan.com/drugdiscovery To learn more about Novartis, go to www.novartis.com
“One of the most difficult aspects of the project was building this level of flexibility into a fully validated system that meets our stringent quality assurance criteria and complies with GLP/GCP regulations. With new assay technologies constantly coming onto the market, it was important to have a system that could adapt to changes in our workflow, and Tecan’s application knowledge
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Genomics solutions for marine biology A Freedom EVO® platform is helping scientists at Italy’s Stazione Zoologica Anton Dohrn, one of the world’s top marine biology and ecology research institutions, to provide a range of automated genomics services for marine biologists.
“With so much flexibility available to us, there are a lot of potential applications.”
The Molecular Biology and Bioinformatics Unit at the Stazione Zoologica Anton Dohrn of Naples provides a range of research services – including DNA sequencing, qPCR, NGS, technical consultations and training in molecular biology – as well as on-demand protocol development. As part of the European Marine Biological Resource Centre (EMBRC), it is a core laboratory focused on developing and optimizing a variety of protocols to enable the study of new marine organisms. Automation has an important role to play, offering maximum flexibility and enabling the department to provide rapid, high quality sequencing, as Dr Elio Biffali explained: “Our department supports researchers at Stazione Zoologica and external organizations, providing routine services such as Sanger and next generation sequencing, real-time PCR and mini-preparations of plasmidic DNA. Working in the marine environment brings its own challenges, but this is also what makes it so interesting, as the extent of biodiversity means that there is a wide assortment of organisms for us to work on. Marine biologists do not use typical model organisms – such as mice or rats – but algae or other marine organisms, which are more difficult to work with. For example, the marine environment makes it difficult to access and retrieve samples or organisms, reducing the amount of material we have to work with. In addition, the composition of the animal tissues has evolved to accommodate the salty environment – some algae, for example, produce foam – and any residual salt from the ocean must be removed before the DNA or RNA can be extracted and purified. There is also a lack of available reference material. Nothing is standardized in such a biodiverse environment.” The unit has invested in a Freedom EVO 200 liquid handling platform to help with this work, allowing it to automate protocols for more complex and high throughput projects. The system is equipped with dual liquid handling arms – a MultiChannel Arm™ 384 for pipetting in 96- or 384-well formats, and
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an eight-channel Liquid Handling Arm with four large and four low volume syringes – as well as a gripper tool for manipulating microplates on the workdeck. In addition, the system has two MIO™ incubators, a Te-Shake™ shaker, a Te-VacS™ vacuum manifold, hotels and all the tools necessary for automated in situ hybridization, as well as a circulating water bath with a refrigeration system. The platform also has the capability to perform automated electrophoresis protocols using the E-Gel® system (Life Technologies). “Our group’s philosophy is to ensure that we have – and maintain – the highest flexibility possible,” Elio continued. “When we came to replace our existing system, which is no longer supported, we looked for a platform that combined the maximum possible flexibility with speed and, obviously, high quality. We spoke to various manufacturers, eventually choosing the Freedom EVO because it offered the best solution in terms of costs, throughput and flexibility. Moreover, this system provides the best technology for our automated in situ hybridization protocols and can be freely configured to meet our needs. This allows us to develop high quality, flexible protocols.” Dr Marco Borra, EMBRC-IT liaison officer, added: “The Freedom EVO is very intuitive, and it is really easy to reconfigure the system and adjust the workdeck layout as our needs change. The Freedom EVOware® software is quite open and user friendly, and is everything that we asked for. We received excellent on-site training from Tecan and, as we familiarized ourselves with the system, learnt to create and optimize new protocols. This has enabled us to develop automated protocols for the kits we most commonly use, even though most are intended to be performed manually.” “With the Freedom EVO, we can alternate between different high throughput screening protocols, automatically switching between 96- and 384-well formats very rapidly
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during a run, which significantly increases our throughput compared to our previous workstation. We’ve had the system for about a year now, and we are transferring all our day-to-day protocols – real-time PCR, Sanger sequencing, mini-preparation and gDNA extraction – onto the Freedom EVO. In the near future, we also plan to implement protocols for our next generation sequencing, including library preparation and the different steps of library enrichment. Eventually, we intend to run all our PCR sequencing reactions on the Freedom EVO, and hope to introduce cherry-picking for library screening. With so much flexibility available to us, there are a lot of potential applications,” Marco concluded. To find out more about Tecan’s genomics solutions, visit www.tecan.com/genomics To find out more about the Molecular Biology and Bioinformatics Unit at Stazione Zoologica Anton Dohrn, visit www.szn.it and www.sbmweb.it
Elio Biffali and Marco Borra from the Molecular Biology and Bioinformatics Unit
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Driving oncology drug development forward The HP D300 Digital Dispenser has enabled Translational Drug Development in the USA to automate its workflows, increasing sample throughput and the number of cell line panels handled, as well as enhancing workflow efficiency.
Translational Drug Development (TD2), based in Scottsdale, Arizona, is a contract research organization offering specialist services to biotech and pharma companies, helping to ensure that cancer patients have rapid access to the most up-to-date oncology treatments. To decrease the time spent manually pipetting and improve efficiency, the company invested in an HP D300 Digital Dispenser, as Paul Gonzales, vice president of nonclinical operations, explained: “TD2 specializes in oncology drug development, providing a wide range of services from in vitro and cell culture assays through Phase II clinical development. We run proof of concept studies that our clients may not have the capabilities to perform, using our expertise to develop strategies to establish and fully explore the activity of a compound. The project management team is in touch with the client almost daily, enabling practical decisions to be driven by the data. As a smaller organization, we can be flexible, responding rapidly to meet the changing needs of a particular project. This ensures that studies are carried out swiftly, helping to meet regulatory requirements and enable the drug to progress to the clinic more rapidly. Ultimately, it’s all about getting the most effective drugs to the patients in the shortest possible time.”
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Kari Kotlarczyk benefits from the straightforward operation of the HP D300
“The HP D300 was purchased as part of a drive to enhance the efficiency of our processes,” said Paul. “Previously, our protocols were 100 % manual. We wanted to introduce automation to increase our sample throughput and the breadth of our work in respect of the number of screens we perform and the cell line panels we use, enabling us to perform complex combination studies more easily. Automation also has the advantage of relieving staff from the burden of manual pipetting, allowing them to focus on other aspects of the work. We looked at the different options available to us, and the HP D300 was the only instrument that met our needs precisely; when it was demonstrated to us, we fell in love with it right away.”
In vitro research associate Kari Kotlarczyk added: “Before we had the HP D300, I was performing a great many pipetting procedures manually, which was extremely time consuming. The HP D300 is very straightforward and easy to use, reducing the potential for the introduction of manual handling errors, and freeing up time for me to carry out other tasks.” Mario Sepulveda, manager of nonclinical operations, takes up the story: “We’ve had the HP D300 for about a year now, and it is a big improvement on manual processing. Prior to implementing automation, we relied on one research associate to perform all the manual pipetting steps, which wasn’t efficient. Compared to manual workflows,
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the HP D300 saves us so much time. If we need to investigate a large number of compounds, we can do a combination rather than a single agent, which is far quicker; whereas before we could handle 20 cell line panels at the most, we can now do up to 60 panels in the same time frame.” “Accuracy and precision are crucial,” explained Mario. “The HP D300 dispenses extremely small volumes with very good accuracy and precision. We now use much smaller quantities of our stocks than before, which is better for our clients. When you are carrying out drug development studies, the amount of compound available can be limited and is very precious; it is important
not to use more than necessary. Most of the time we are preparing drug dose-response curves to determine the IC50 value, selecting the cell lines in which the specific compound of interest is active to try to move forward to in vivo testing. This can involve many dilution steps, and the elimination of manual pipetting really improves the efficiency of these experiments, giving us total confidence in the results. Although it is not common, if a dose-response curve does produce an unexpected result, we can be 100 % sure that the dilutions are correct and that the discrepancy is not due to a manual error. This allows us to focus on probable compound-related causes.”
“…the elimination of manual pipetting really improves the efficiency of these experiments, giving us total confidence in the results.” Mario continued: “The software is simple to use and there is so much you can do with it. We can randomize plate layouts and perform even the more difficult dilutions with ease, allowing us to really define the dose-response curve and establish the IC50 or EC50 value. For drug synergy studies, we use a checkerboard design. The limitations of manual processing have been considerably reduced, allowing us to thoroughly interrogate double, and occasionally triple, combinations in a very efficient and accurate manner. The HP D300 really works out.” Paul concluded: “Where we really make savings is on personnel efficiency, avoiding the need to take on additional staff to accommodate a specific study. That, coupled with data integrity, is the most important thing to me. It’s a no brainer.” To find out more about the HP D300, visit www.tecan.com/digitaltitration To find out more about Translational Drug Development, visit www.td2inc.com
The TD2 team. Left to right: Paul Gonzales, Caileen Walker, Kari Kotlarczyk and Mario Sepulveda
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Developing the fuels of the future Extended periods of walkaway operation are essential to the work carried out by Spanish energy producer Repsol, which relies on two Freedom EVO® platforms and Tecan’s 350 µl nested liquid handling disposable tips for its research into alternative energy sources.
Scientists at the Madrid Technology Center of global energy company Repsol are engaged in research to develop fossil fuel alternatives, such as advanced biofuels. More than 400 researchers and scientists work together at the center to improve the value of the energy chain through technological innovation, contributing to a safer, smarter and cleaner energy model. To help drive this research forward, the company has invested in two Freedom EVO platforms for colony-picking and enzymatic assays, relying on nested disposable tips to achieve the extended walkaway times required. Jose Miguel Seoane, a researcher in
the biotechnology department, explained: “The investigation of new biological applications involves screening large numbers of mutants of different microorganisms, enabling the best performing candidates to be selected for further studies. As we test thousands of different mutations, automated high throughput screening is the key to our success.” Jose continued: “We start by targeting a particular molecule, using standard microbiological methods to produce it in the laboratory. At the same time, our computational team is performing
The biotechnology department’s Freedom EVO platforms are central to its research
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thousands of calculations, exploring all the possible mutations and modifications of the selected enzyme or biological pathway to establish what is likely to generate the greatest possible yield. These results are implemented in the laboratory, creating thousands of clones which must be explored to see whether they perform as expected. In this situation, high throughput screening is essential, allowing us to identify the best candidates to take forward. Eventually, after a few cycles of directed evolution – protein engineering – and further screening, we can progress towards our goal of producing a commercially viable product.”
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“Automation is vital for this work; the clones are isolated on a Freedom EVO workstation equipped with a colony-picker (Pickolo™, SciRobotics), Robotic Manipulator (RoMa) and Liquid Handling (LiHa) Arms, 350 µl Tecan Sterile nested LiHa disposable tips, a plate sealer and a carousel for Petri dishes (SciRobotics). Petri dishes containing the colonies are stored in the carousel and the mutants harvested using the colonypicker. After transfer to a microplate, overnight incubation allows the mutants to be grown separately in individual plate wells prior to performing enzymatic assays on a second Freedom EVO system. This instrument is equipped with a RoMa Arm, a MultiChannel Arm™ 96, 200 µl Tecan Sterile disposable tips – standard and nested – three Teleshakes (INHECO), a Te-VacS™, a 10-plate hotel, a Tecan Carousel and an Infinite® 200 PRO microplate reader, as well as Magellan™ data analysis software. The target protein is extracted, and enzymatic assays are carried out to determine whether the modification introduced makes a difference or not.”
Nested 350 µl LiHa disposable tips – passive stack of five layers
“We looked at systems from several companies before choosing the Freedom EVO,” said Jose. “I had used Tecan systems in my previous employment and had been very happy with them, and the Freedom EVO is very well tested with a proven track record. The company also offers excellent support, and had the advantage of being able to provide a colony-picking workstation and a high throughput platform. Of particular interest to us was the availability of nested LiHa disposable tips. We generate up to 10,000 colonies per week, and long periods of walkaway automation are absolutely essential. It makes no sense to invest in automation if you need to manually place tips on the workdeck every 20 minutes or so; nested tips are indispensable. The release of 350 µl nested LiHa disposable tips, with a novel design incorporating a frame between each tray, was an important development. The trays fit perfectly and the stack remains in perfect alignment, eliminating tip pick-up failures. This makes the workflow more robust and efficient, as well as providing the long walkaway times that we need.” “We have had the Freedom EVO systems for about a year now, and choosing them was a good decision. It would be almost impossible and extremely complicated to do this work manually. Automation is much more precise,
“Of particular interest to us was the availability of nested LiHa disposable tips. We generate up to 10,000 colonies per week, and long periods of walkaway automation are absolutely essential.” giving us total control of the process, with traceability provided by barcodes. We have developed robust Freedom EVOware® scripts that are straightforward to use and, by running the enzymatic assays overnight, gain 10 to 12 hours as the samples are ready for us to use in the morning. The support from Tecan is excellent; the field engineers know the systems well and are very responsive, which we really appreciate as a project can be set back weeks, or even months, if the support is not there. It was a pleasure to work with the company,” Jose concluded. To find out more about Tecan’s nested liquid handling disposable tips, visit www.tecan.com/lihaditi To learn more about the Repsol Technology Center, go to www.repsol.com
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Stressing the importance of quality assays The Department of Psychology at TU Dresden is taking advantage of the quality and reproducibility of various IBL International immunoassays – including the Cortisol Saliva Luminescence Immunoassay – to investigate the link between steroid hormones and stress.
Researchers in the Department of Psychology at Germany’s Technische Universität Dresden (TU Dresden) have been at the forefront of stress research for over 30 years. The department uses a broad range of techniques – from ‘question and answer’ and paper-based testing through to brain imaging and biochemical assays – to investigate the effects of both acute and chronic stress on the body. Professor Clemens Kirschbaum explained: “Psychology has come a long way since the days of simple ‘pencil and paper’ testing, with many research centers now routinely employing electroencephalography (EEG) and functional magnetic resonance imaging (fMRI), but there are still only a limited number of wet labs looking into the biochemical changes associated with psychological profiling.” “Steroid hormones are a key area of interest for us; any deviation from the physiological ‘normal’ levels of biomarkers such as cortisol can be used as indicators of acute or chronic stress conditions. However, unlike most clinical biomarker testing – which uses
blood, urine or cerebrospinal fluid – we need non-invasive testing methods to avoid skewing results, and so have developed a range of techniques based on saliva or hair extract analysis.” “The measurement of cortisol in saliva is one of our most commonly performed assays, and we first developed a microplatebased chemiluminescent immunoassay for this application in 1986. As study sizes grew bigger and bigger – many individual studies in this area now comprise more than 10,000 individual samples – our in-house assay became impractical in terms of the labor involved in hand-coating the plates, as well as the batch-to-batch reproducibility and long-term stability. It was clear that commercially produced assay plates, with the associated quality control measures, were a more practical option. We switched to IBL International’s assay in 2001, and have been very happy with our decision ever since.” “We are quite active in supporting laboratories and research groups around
Members of the Department of Psychology wet lab team with one of the department’s Tecan liquid handling workstations
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“We switched to IBL International’s reagents in 2001, and have been very happy with our decision ever since.” the world, receiving samples from across Europe, almost every state in the USA, Australia, New Zealand, Japan, Peru and many other countries, and perform many thousands of tests a year for the cortisol assay alone. As most of these groups are academic researchers, the cost of testing is an important consideration, especially with so many samples per study. Combining the IBL assay kits with our Tecan liquid handling workstations certainly helps us to efficiently process the samples in a time- and costeffective manner, enabling us to check every result to ensure high quality datasets for our colleagues and collaborators.” “Unlike a contract testing laboratory, we have a real interest in our partners’ research. With over 25 years of experience performing this type of assay, as well as designing and running studies ourselves, our collaborators trust us to provide a high quality service. Offering this testing service also gives us unprecedented access to datasets from studies around the world, all processed in exactly the same way. For these reasons, our reputation is very important to us, making it essential to have a long-term partner we can trust and work with to expand and improve the quality of our testing. IBL International is exactly this, and the company has always been very responsive to our needs. We have a very good relationship with the Hamburg team; they are always keen to help us as quickly as they can and we regularly share data and ideas on the development of new or improved assays.”
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“Alongside our immunoassay testing, techniques such as LC-MS are becoming increasingly prevalent for biomarker analysis, and we are also using our Tecan liquid handling workstations to automate sample preparation for some of these other modalities, for example, we are now performing liquid-liquid extractions on the platforms. IBL International becoming part of the Tecan Group offers the potential for further collaborations, allowing us to further benefit from our long-term relationship and contribute towards future developments in this exciting area.” To find out more about IBL International’s range of saliva diagnostic kits, visit www.ibl-international.com/en/salivadiagnostics To learn more about the Department of Psychology at TU Dresden, go to http://biopsychologie.tu-dresden.de
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A simple(x) approach to bioprocessing UCL’s Advanced Centre for Biochemical Engineering is harnessing the flexible automation capabilities of its Freedom EVO® workstations to explore novel bioprocessing applications, with a focus on the production of ‘next generation’ medicines including protein-derived therapeutics, vaccines and cells for therapy. Combining the Freedom EVO platform with devices such as Atoll’s MediaScout® RoboColumns®, researchers are able to generate high quality, scalable data that minimizes the cost and time required to optimize new bench-scale processes.
University College London’s Department for Biochemical Engineering is a world leader in the development of novel bioprocesses, focusing on macromolecular and cellular processing techniques for industrial and biomedical applications. The department’s Advanced Centre for Biochemical Engineering (ACBE) is an interdisciplinary facility that coordinates research collaborations with other UCL departments, as well as academic institutions and industrial partners around the globe. The ACBE combines cutting-edge laboratory
automation with the latest industrial process development tools to accelerate its research, as Professor Ajoy Velayudhan explained: “Automation is now an integral part of processing biological materials, even at the research and development level, and we currently have six Freedom EVO platforms within the department. The majority of these instruments are set up for generic liquid handling activities, providing convenient and flexible walkaway automation to meet the needs of individual projects. We have a number of modules that
Members of the ACBE team with one of the department’s Freedom EVOs
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can be installed or removed as required, allowing us to do everything from simple biochemical assays to complex cell-based studies. For example, we can reproducibly expand and differentiate stem cells, as reported in an earlier edition of the Tecan Journal (Issue 3, 2009).” “One of our areas of particular interest is the development of quantitative models for whole bioprocesses, looking at how the individual steps fit together to yield the final product. These bioprocesses generally
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consist of 10 to 15 individual steps, each of which can influence processes downstream. We are developing methods to evaluate the ‘cross-talk’ between these operations, requiring large quantities of data. This data generation is extremely tedious, and would be virtually impossible to achieve manually. High throughput automation techniques are therefore vital to generate the rich datasets necessary to extrapolate meaningful results.” Another topic currently under investigation is the optimization of polishing steps for the final purification and clean-up of therapeutic biological products. The ACBE team is taking advantage of the Freedom EVO’s liquid displacement and Te-Chrom™ module to provide walkaway processing of Atoll’s MediaScout RoboColumns. Dr Spyridon Konstantinidis explained: “We use the RoboColumns to assess different operating conditions for preparative chromatography applications, optimizing objective functions such as purity, yield, throughput and productivity of protein purification. Combining these columns with the Freedom EVO workstation is very advantageous for our work, allowing parallelization of screening and enabling much more rapid progress than would be possible using traditional benchscale testing. We have found the results of experiments conducted using RoboColumns to be scalable, making this a useful tool for production-scale process optimization for the generation of the extremely pure biomolecules needed for therapeutic use.”
A major advantage of this set-up is the small amount of material required for process optimization. This can often be a significant obstacle, due to the high costs of producing material for screening studies. To further reduce consumption of biological materials, the ACBE has developed a modified simplex optimization method which allows rapid development of processes without the drawbacks of traditional Design of Experiment (DoE) approaches. Ajoy continued: “DoE techniques are excellent for late-stage commercial development, where robustness around an established set of operating parameters needs to be demonstrated. However, for the type of novel processes we usually study, DOE can be laborious and inflexible in finding effective operating parameters. Experienced researchers generally prefer to analyze data on-the-fly, allowing them to rapidly progress to the next stage of optimization without needing to run a large number of unnecessary ‘peripheral’ experiments. A more effective approach to early process development uses the classical simplex method. We have generalized this method to account for both continuous and discrete variables, making it much more useful for bioprocess development. It is highly adaptable, saves time and material, and is closer to the intuitive way experienced scientists do process development. However, this requires a very dynamic approach to automation, which our Freedom EVO platforms provide.”
“This requires a very dynamic approach to automation, which our Freedom EVO platforms provide.” Spyridon added: “The performance of Tecan systems is well documented in terms of accuracy and precision, but the flexibility of the Freedom EVO instruments is even more important for us. The workstations are also quite easy to use, allowing even relatively inexperienced postgraduate students to get processes up and running very quickly with little training. At the other end of the spectrum, the open nature of the system allows us to compile our own executable applications, which can then be integrated with Freedom EVOware® to create automation scripts on-the-fly. As long as you set it up correctly, you know you’re going to get quality data – it’s very reliable, and the support we’ve received from Tecan has been brilliant.” To find out more about Tecan’s bioprocessing solutions, visit www.tecan.com/bioprocessing To learn more about University College London’s Department for Biochemical Engineering, go to www.ucl.ac.uk/biochemeng
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Freedom to evolve Researchers at the Complutense University of Madrid are using directed evolution techniques to develop new enzymes for a variety of bioprocessing applications. Taking advantage of the walkaway liquid handling capabilities of a Freedom EVO® 75 workstation, the university’s Enzyme Biotechnology Group – headed by Dr Isabel de la Mata – has been able to investigate ~4,000 Rhodococcus mutants for penicillin acylase and N-acyl homoserine lactone acylase activity, conducting over 190,000 assays in the primary screen alone. increasingly prevalent in nosocomial pathogens. We are also using this approach to develop novel antifungal agents, such as echinocandins, as there are currently very few effective therapeutic antifungals.”
The Enzyme Biotechnology Group – part of the Biochemistry and Molecular Biology Department I – at the Complutense University of Madrid, Spain, has been investigating industrial applications of enzymes for over 30 years. One of the group’s current research areas is the investigation of enzymes with potential applications in the synthesis of therapeutics, as Dr de la Mata explained: “We are interested in enzymes with novel activities for use in the production of semi-synthetic antibacterial agents, such as β-lactam antibiotics, to overcome the resistance mechanisms which are becoming
“We use directed evolution techniques to look for mutations which modify the substrate specificity or catalytic activity of the target enzyme, as well as its stability in differing environmental conditions, such as variations in pH or temperature or presence of organic solvents. The success of these molecular evolution methods depends on the ability to investigate large numbers of clones, and so we needed a stable micro-organism expression system suitable for high throughput screening. Unfortunately, many of the enzymes we study cannot be expressed in the commonly used E. coli models, and so we have had to develop an alternative system
Left to right: Rodrigo Velasco, Miguel Arroyo and Isabel de la Mata with the group’s Freedom EVO 75
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using Rhodococcus sp. strain T104. Rhodococcus is far more difficult to work with than E. coli but, using our Freedom EVO 75 platform, we have been able to develop a robust high throughput assay for screening the mutants in a 96-well microplate format.” The group’s Freedom EVO 75 platform is equipped with an 8 Plus 1 Access™ arm, a MIO™ incubator and temperature-controlled carriers for assay plates and reagents, allowing walkaway processing and reproducible results. Rodrigo Velasco, a PhD student in the group, commented: "Thanks to the throughput capabilities of the Freedom EVO platform, each sample could be simultaneously assayed under several operational conditions, which would be very complex and difficult to achieve manually. This provides invaluable information to help us design the downstream stages of the research.”
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Dr de la Mata continued: “Using the Tecan platform we have been able to develop a sensitive, high throughput colorimetric screening method to identify, for example, new acylase specificities. In the first round of screening we investigated over 4,000 individual clones, each tested in duplicate, looking at a variety of conditions: two pH values, two temperatures and seven different substrates. This experimental design resulted in over 190,000 individual assays, which would have been impossible to perform manually, but using the Freedom EVO we were able to process the samples faster than we could analyze the results! The primary screen gave us a number of promising candidates with characteristics of interest for technological applications, which were then taken forward to create a second generation of mutants for screening. This second round of testing consisted of around 27,000 assays, and yielded several highly efficient biocatalysts for further investigation, all within two years of beginning the project.” “We have been very happy with the performance of the Freedom EVO workstation and the assistance we have received from Tecan. This type of project simply wouldn’t have been possible to perform manually, and the technical support has been excellent,” Dr de la Mata concluded. To find out more about Tecan’s Freedom EVO 75, visit www.tecan.com/freedomevo75 To learn more about the Complutense University of Madrid, go to www.ucm.es
“We were able to process the samples faster than we could analyze the results!”
Werner Hälg, Corporate IP Coordinator
Leading the debate When people hear the term ‘intellectual property’ (IP), they usually only think of patents. However, IP also includes trademarks, design rights, internet domain names, copyrights and trade secrets, all of which can play an important role in a company’s ability to meet the needs of customers. As a leading manufacturer of laboratory automation technologies, Tecan constantly strives to offer faster, more robust and more efficient solutions to address the specific needs of the life sciences market. Innovation is a crucial aspect of what we do as a company, and supporting this with a strong IP portfolio is an important part of ensuring we offer our customers the very best technical solutions and support. Although the end user may not be aware of it, every Tecan system is packed with unique, proprietary technologies which help to ensure more efficient and reliable performance, from the Spark™ 10M’s Humidity Cassette – which minimizes evaporation during assays – to Adaptive Signal Technology™, a new generation of liquid level sensing introduced with the Fluent™ workstation. The software and its features are also protected by copyright law. Trademarks and registered designs are another important aspect of IP. These don’t just stop another supplier from using a product’s name or ‘look and feel’, they are an important way of offering the customer a ‘guarantee’ that they are buying a genuine Tecan product, backed by our liquid handling expertise and application knowledge. This innovation, quality and integrity is especially important for our Partnering Business. Our OEM customers rely on Tecan to develop and build high quality, effective automation solutions that offer troublefree operation and high quality results. This helps to ensure a positive experience for end users and to protect our partners’ reputations and market positions. To let us know your thoughts, contact us at hello@tecan.com Find out more at www.tecan.com/linkedin TALK TO TECAN TECAN JOURNAL 2/2015
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Atlanta, USA
26 – 30 July 2015
ASHG The American Society of Human Genetics 2015
Baltimore, USA
06 – 10 Oct 2015
SOFT Society of Forensic Toxicologists 2015
Atlanta, USA
18 – 23 Oct 2015
BioProcess International Conference & Exhibition
Boston, USA
26 – 29 Oct 2015
Tsukuba, Japan
01 – 03 July 2015
43 Annual General Meeting of the Australian Society for Microbiology
Canberra, Australia
12 – 15 July 2015
25 Australian and New Zealand Society for Mass Spectrometry/6 Asia Oceania Mass Spectrometry Conference
Brisbane, Australia
19 – 22 July 2015
39th Human Genetics Society of Australia Annual Scientific Meeting
Perth, Australia
08 – 11 Aug 2015
QMB Queenstown Molecular Biology Meeting 2015
Queenstown, New Zealand
22 – 28 Aug 2015
25th Annual Combined Biological Sciences Meeting
Perth, Australia
28 Aug 2015
Asia and Pacific NGS Field 4th Meeting rd
th
th
JASIS Japan Analytical & Scientific Instruments Show 2015
Chiba, Japan
02 – 04 Sept 2015
2015 National Laboratory Medicine Conference
Nanjing, China
09 – 12 Sept 2015
JACLaS Japanese Association of Clinical Laboratory Systems EXPO 2015
Yokohama, Japan
08 – 10 Oct 2015
The 16 Beijing Conference and Exhibition on Instrumental Analysis
Beijing, China
27 – 30 Oct 2015
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Paris, France
22 – 24 June 2015
MSACL EU Mass Spectrometry Applications to the Clinical Lab
Salzburg, Austria
09 – 11 Sept 2015
MipTec 2015
Basel, Switzerland
22 – 24 Sept 2015
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Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Günter Bolzern/Zürich www.bolzern.tv Editor in Chief: Tecan Trading AG, Antonietta Allocca Editor: kdm/UK www.kdm-communications.com Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, journal@tecan.com, www.tecan.com To register for the Tecan Journal please go to www.tecan.com/journal © 2015 Tecan Trading AG, Switzerland, all rights reserved.
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EVENTS TECAN JOURNAL 2/2015
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