A fresh approach to clean water

page 30-31

Edition 2/2012

Tecan JournalLife Sciences and Partnering Business

High stakes laboratory automationpages 12-13

A real winner for celiac patientspages 20-21

The right tools to drive drug discoverypages 28-29

2 CEO WELCOME TECAN JOURNAL 2/2012

Welcome to this issue of the Tecan Journal

As I look back over my long career in the life sciences industry, I am proud to take on the role of acting CEO of Tecan until my permanent successor joins the Company later in the year. This transition comes at an exciting time for the Company, following the launch of some innovative new products at this year’s SLAS Conference and Exhibition in San Diego, California. The exhibition heralded success for Tecan, with the new HP D300 Digital Dispenser one of just four products to receive the prestigious New Product Award Designation, and the forthcoming launch of the Air LiHa (Air displacement pipetting arm) for the Freedom EVO® generating a lot of interest. We are also very excited about the expansion of our business in Asia and Australia and, in this issue, you will find information on the recent opening of our new facilities in Shanghai and Melbourne.

This growth of our business is being underpinned by our global quality program, and we have details of how the latest developments of this program are helping to further strengthen our support for our customers around the world.

Also in this issue are details of recent updates to the Gas Control Module (GCM™) for the Infinite® 200 PRO microplate reader, together with the results of the 2011 Tecan Award for detection instruments. There is also a variety of articles about how our customers are using Tecan equipment for a diverse range of tasks – from drug testing for the horse racing industry to the purification of drinking water – relying on our automation expertise and reputation for quality to help further their life science applications.

We hope you enjoy the issue.

Gérard Vaillant, Acting CEO

Wel

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3CONTENTS TECAN JOURNAL 2/2012

2 CEO Welcome

4 Simplifying QC for Freedom EVO workstations

4 Tecan ends support of GENios™ plate reader

5 Enhanced Gas Control Module (GCM) now offers simultaneous control of CO2 and O2

6 Tecan Award 2011 winner announced

7 China Strategy Project provides new investment in Asian market

8 Direct Digital Titration triumphs at SLAS2012

8 2012 Tecan Symposium – mass spectrometry

9 Customers celebrate the launch of Tecan Australia

10-11 Creating a Rosetta Stone for certification

12-13 High stakes laboratory automation

14-15 Developing new, efficient immunodiagnostic technologies

16-17 Finding the right candidate

18-19 Caring for staff and patients; benefits all round with the FE500™ and a Te-Care™ Service Contract

20-21 A real winner for celiac patients

22-23 Capturing the ‘dynamics’ of automated cell culturing

24-25 A powerful combination for qPCR

26-27 Automation aids peptide biomarker discovery

28-29 The right tools to drive drug discovery

30-31 A fresh approach to clean water

31 Leading the debate

32 EventsCont

ents

page 8

pages 30-31

pages 24-25

page 5

A fresh approach to clean water

2012 Tecan Symposium

4 PRODUCT NEWS TECAN JOURNAL 2/2012

Simplifying QC for Freedom EVO® workstationsTecan has partnered with Artel – the worldwide leader in liquid handling quality assurance – to offer the QC Kit, a fully integrated pipetting performance verification solution for Freedom EVO liquid handling workstations.

The QC Kit will take advantage of Artel’s proprietary Ratiometric Photometry™ technology to provide clinical, pharmaceutical and forensic laboratories with a simple and reliable way to meet increasing quality and regulatory demands for performance verification.

The QC Kit includes an Infinite® F50 or Infinite 200 PRO microplate reader, a calibrator plate, a microplate shaker, a barcode scanner and a Freedom EVOware® software add-on. Characterized verification microplates and color dye solutions will be available for ordering directly from Tecan. All color dye solutions are traceable to international reference standards, allowing direct comparison of precision and accuracy performance data between different sites and instruments. Liquid handling performance results can be obtained automatically in just 10 to 30 minutes – depending on the workstation configuration – without the need for sample preparation, standard curve generation or a carefully controlled testing environment.

QC Kit dye solution

Artel is a registered trademark and Ratiometric Photometry is a trademark of Artel.

The QC Kit is under development and will be available in the near future.

To find out more about Tecan’s QC Kit, visit www.tecan.com/qckit

Tecan is happy that thousands of satisfied customers have been using this robust instrument for decades, but it has now been replaced by a new generation of multimode microplate readers – the Infinite 200 PRO series – offering:

• Fastermeasurementofmoresamplesinsmaller volumes

• Measurementofsmallvolumesamplesin96- and 384-well microplates

• Predefined‘one-click’DNAmeasurementswith user-friendly i-control™ software

• Automaticcalculationof260/280 purity ratios

• Measurementofupto162µlDNA drops in one run, with the patented NanoQuant Plate™

• Eliminationoffuturecompatibilityproblems with computers and printers

Tecan ends support for GENios™ plate readerAfter its launch in 1999, the GENios plate reader became one of the most popular plate readers in the life science community. But, even the best things must come to an end, and Tecan has announced it is ending support for this well established, but past its prime, microplate reader.

Talk to your Tecan sales representative to see how we can help you switch to a new generation of microplate readers, or visit www.tecan.com/dna-reader to learn more.

5PRODUCT NEWS TECAN JOURNAL 2/2012 5PRODUCT NEWS TECAN JOURNAL 2/2012

Enhanced Gas Control Module (GCM™) now offers simultaneous control of CO2 and O2

Launched in 2010, the patent pending Gas Control Module (GCM) for the Infinite 200 PRO multimode reader was a breakthrough for cell -based experiments, offering precise regulation of oxygen or carbon dioxide levels within the reader’s detection chamber for the first time. This powerful module has now been further enhanced, allowing simultaneous control of O2 and CO2 concentrations and providing rigorous environmental control for an even wider range of cell biology applications.

The enhanced GCM uses improved sensor technology to increase measurement accuracy and minimize cross-interference for oxygen measurements. Combined with redesigned, independent gas inlets, this offers precise, independent control of both O2 and CO2 levels within the reader, helping to further stabilize culture conditions for more consistent cell growth. The greater

sensitivity and precision offered by this set-up also allows the instrument to automatically compensate for variations in the atmospheric partial pressure of CO2 using the module’s unique altitude correction function, which is accessed via an intuitive new interface.

Process security has also been enhanced, providing rapid detection of changes in gas pressure or flow to identify if the target concentration will not be reached or deviates significantly during incubation, with both audible and visible warnings to alert the user. A new version of the Infinite 200 PRO’s i -control software will also soon be available to take full advantage of these enhancements, providing a real-time online display of gas concentrations on your PC during operation, and generating a log of gas concentrations throughout the experiment. This data will also be added to the results file alongside each data point, ensuring

comprehensive process monitoring and maximum data integrity.

These new and improved features ensure greater biological relevance for a wide range of studies, extending the experimental window for microplate-based investigations of anaerobic or facultative anaerobic bacteria, and allowing close replication of hypoxic or physiological conditions for eukaryotic cells. Together with the Infinite 200 PRO’s cell biology oriented functions – including Optimal Cell Reading (OR), linear and orbital shaking, advanced temperature control and automated z-focusing – this comprehensive solution sets new standards for cell-based assays.

To find out more on the GCM for Tecan’s Infinite 200 PRO, visit www.tecan.com/gcm

6 CORPORaTE NEWS TECAN JOURNAL 2/2012

Tecan Award 2011 winner announcedEstablished in 2010 to celebrate the innovation and ingenuity of Tecan’s loyal customers, the Tecan Award highlights some impressive and imaginative uses for the Company’s detection instruments. We are pleased to announce that the 2011 award has been won by Svenja Kristina Holle from the University Hospital of Muenster (UHM), Germany, for her work with an Infinite® 200 microplate reader. Svenja and the UHM team have developed an ex vivo method for investigating the dynamics of organic cation transport in proximal tubules of the kidney. This innovative optical method could be adapted to almost any kind of freshly isolated biological transport system using a suitable fluorescent substrate, offering an exciting new technique to study membrane transport.

Second place in the 2011 competition went to Michael Bunge from the Institute of Applied Microbiology at Giessen University, Germany, who is using an Infinite M200 to monitor the effects of a metallic nanoparticle-based approach to eliminating the growth of potential pathogenic microorganisms in drinking water (featured on page 30 of this issue). And a close third place went to María Isabel Pividori from the Universitat Autònoma de Barcelona (UAB), Spain, for developing a novel method for the quantification of gliadin in gluten-free foods using a Sunrise™ reader (see page 20 for more information).

The 2011 award will be presented to Svenja during a tour of Tecan’s Detection Headquarters in Grödig, Austria, as part of her prize of a trip to the romantic city of Salzburg.

To find out more about the Tecan Award and details of the winning entries, visit www.tecan.com/award

Second Place: Michael Bunge (third from left) from the Institute of Applied Microbiology at Giessen University, Germany, see feature page 30

Third Place: María Isabel Pividori (far right) from the Universitat Autònoma de Barcelona (UAB), Spain, see feature page 20

China Strategy Project provides new investment in Asian market

When Tecan established its Beijing representative office in 2004, customer service and support were already important components for the development of our business in China. This was strengthened in 2008 by the creation of Tecan’s Asia Pacific Headquarters, with local service manager Hua Zhang and Johann Prossinger, originally from Tecan Austria, tasked with developing the local service capabilities. The launch of our China Strategy Project last year has seen further development of our local service operation, including the creation of a new service and training center in Shanghai, and the roll out of a local helpdesk service, which has been a winning practice within the Tecan Group for many years.

The Asia Training and Service Center reflects our strong commitment and continuing investment in the Asian market, and houses a comprehensive sales and support organization, complete with customer training and demonstration facilities and local application development, to provide rapid support for customers’ needs. Inaugurated on the 15th of March this year, the opening ceremony included a scientific seminar and tour of the new facility, and was attended by over 60 distinguished guests from Tecan’s healthcare, industry and academia customers in Asia. The seminar’s theme – Laboratory automation: Advancing science and improving healthcare – attracted

key opinion leaders in public healthcare policy, diagnostics and pharmaceutical development, and was followed by an open forum discussing current and future laboratory automation applications in drug discovery, translational medicine, and clinical testing and research.

To complement the new service and training facilities, we are also establishing a local helpdesk in China for the first time. The new China helpdesk team is comprised of two dedicated engineers and two back-up engineers from the customer service team, led by experienced Tecan field engineer Billy Gu. Billy and his team have worked closely with a group of experts from Tecan US – including Vice President of Service and Consumables for Tecan US, Craig Williamson – to set up this service, combining local knowledge with established practices and many years of experience to create the best solution for customers. The US and China teams worked shoulder to shoulder to implement and streamline the service – from protocols to system set-up – in just two weeks, aligning the China helpdesk with other departments and business units before the service went live on the 9th of March.

While still at the very beginning of its operations, Billy is totally convinced that all the field experience he and his team have gained during their many years of service

at Tecan is now finding a very effective platform to help all our customers in China. The helpdesk will not only help to solve customers’ problems in a faster and more efficient manner, it will also significantly improve the overall customer satisfaction level in China, which, needless to say, is our ultimate goal.

Raymond Chan, Tecan Head of China and South East Asia, commented: “The rapid development of our business in Asia meant that our former facilities could no longer meet our needs. Establishment of the Asia Training and Service Center, along with the local helpdesk, will extend the level of support we can offer our customers, improving our overall service and strengthening our business in the region.”

To contact the China helpdesk, call the toll-free hotline on 400 821 3888, or e-mail helpdesk cn@tecan.com

Guests at the opening of the new center enjoyed an opportunity to meet fellow Tecan users

The inauguration ceremony brought together many of Tecan’s customers in Asia for a scientific seminar and tour of the new facility

Billy Gu, Team Leader of Tecan China Helpdesk

The Asia Training and Service Center provides a local presence

7CORPORaTE NEWS TECAN JOURNAL 2/2012

An intensive selection process involved an initial assessment phase, where individual judges visited their allocated booths to evaluate and rank each entry before coming together to establish a short list of candidates for the award. Phase two saw the team of judges rating the top nine entries against specific, detailed criteria and, of more than 60 new products launched at SLAS2012, the innovative HP D300 was one of just four chosen to receive this prestigious award.

And it wasn’t just the judges who were impressed by the HP D300. The instrument received widespread interest from delegates, who were impressed by its user-friendly interface and ability to rapidly deliver picoliter to microliter volumes of drug compounds. This latest award is further proof of the convenience and flexibility of Direct Digital Titration and its potential to improve productivity and accelerate the discovery of better drugs.

To find out more about Direct Digital Titration, go to www.tecan.com/digitaltitration

Direct Digital Titration triumphs at SLAS2012The first annual SLAS Conference and Exhibition held in San Diego, California, in February heralded success for Tecan, with the HP D300 Digital Dispenser receiving the SLAS2012 New Product Award Designation.

8 CORPORaTE NEWS TECAN JOURNAL 2/2012

Tecan is proud to announce its fifth Tecan Symposium, to be held this year from the 23rd to the 25th of October in Boston, USA. This year’s topic will be ‘Mass spectrometry – the expanding role in life sciences and diagnostics’ and will include: the application of mass spectrometry (MS) in diagnostics and other fields; the role of mass spectrometry in basic research; the collection, preparation and storage of samples for MS; and novel and emerging MS technologies. As in previous years, attendance at the symposium is limited to a small cohort of scientists in order to encourage plenty of networking opportunities among thought leaders in the area of mass spectrometry.

For more information, visit www.tecan.com/symposium

2012 Tecan Symposium – mass spectrometry

Boston 2012fifth Tecan

Symposium

Boston, October 2012

Jessica Merlino, Tecan Product Manager, and Christie Dudenhoefer, HP Instrument Design Lead, accept the award.

9CORPORaTE NEWS TECAN JOURNAL 2/2012

Customers celebrate the launch of Tecan Australia

Tecan recently invited customers from around Australasia to mark the inauguration of Tecan Australia as a fully owned Tecan subsidiary, with a series of launch events in Sydney, Brisbane and Melbourne. The creation of Tecan Australia reflects Tecan’s uniquely strong commitment to the Australasian market, and will provide customers in Australia, New Zealand and surrounding territories with an even better service, thanks to a new facility in Melbourne and local presence in several major cities.

The launch events gave customers a rare opportunity to discuss their applications and automation issues with Tecan staff and their fellow scientists in a relaxed environment, and proved so successful that Tecan Australia will soon be establishing regular user group meetings to keep these valuable communication channels open.

The Sydney Harbour Bridge offered a spectacular backdrop for the first event

The launch parties brought together Tecan users from around Australasia

The new Melbourne facility offers customers in Australia a local service

The 35th floor dining experience offered a good view of Melbourne

Creating a Rosetta Stone for certification

Following on from the successful ISO certification of our UK, French, German and Benelux organizations, Tecan is now working towards a company-wide matrix certification, to ensure that our organizations around the world are working in the same way, and are collectively striving to improve the services our business offers. To help us achieve this certification, we have chosen to work with TÜV SÜD Product Service, a globally recognized Notified Body with a strong reputation in the life science industry. Lazaros Kostadimas, Tecan’s Global Head of Quality for all Market Units, is

leading the project at a global level and explained: “Tecan has always been centered around the ever-changing needs of its customers, and this project makes sure that we maintain that focus, while continually improving the quality of service we are able to offer. By putting the same processes in place in every country, our global businesses will be far more transparent – allowing us to continually monitor and improve our operations and develop an even stronger customer focus – and we have chosen a process-oriented approach based on ISO 13485 to achieve this.”

Erika Hildebrandt, Quality Manager for Europe, explained: “The first step in this project was to conduct a thorough gap analysis, identifying where there were differences between how we operated in each European market, even in the countries which were already certified. We then held workshops in France and Germany to look at the similarities between local processes, discussing the best ways to handle all aspects of our business for both our customers and for Tecan. This collaborative approach allowed us to bring together the best practices

10 QUaLITY aSSURaNCE TECAN JOURNAL 2/2012

Tecan Systems, Inc.Tecan US, Inc.

Tecan UK Ltd

Tecan Ibérica Instrumentación S.L

Tecan France S.A.S.

11QUaLITY aSSURaNCE TECAN JOURNAL 2/2012

from each country, creating a coherent, standardized process that would best serve the needs of our customers while allowing for the inherent differences between markets. Once this was established, we set up key performance indicators in all countries, allowing us to monitor progress and implement the right corrective actions if necessary. This transparency is really important, helping us to see how we are performing in each country, and how we can improve ourselves at both a local and global level to offer the best service to our customers.”

Lazaros continued: “The first phase of this operation, which included Tecan’s previously certified UK, French, German and Benelux organizations – as well as Italy, Spain, Scandinavia, Switzerland and Austria – has already been completed. Phase two, which is planned for completion in October 2012, will add China and Australia to the scheme, closely followed by the US market unit in 2013 and Japan. For the Quality Team, our focus has been to really keep this project alive, with all Tecan subsidiaries fully aware and in agreement”.

Gerald Schmidt, Head of Market Unit Europe, added: “Everybody involved so far – service managers, sales people, HR, logistics and finance – has worked really hard together as a team to successfully implement this initiative. We have been very impressed with the effort our colleagues in each country have put into it, and the tangible benefits to our customers are already becoming clear.”

Tecan UK Ltd

Tecan Nordic AB

Tecan Australia Pty Ltd

Tecan (Shanghai) Trading Co., Ltd.

Tecan Deutschland GmbH

Tecan Japan Co., Ltd.

Tecan Asia Pte Ltd

Tecan Benelux BVBA

Tecan Ibérica Instrumentación S.L

Tecan France S.A.S. Tecan Austria GmbH

Tecan Software Competence Center GmbH

Tecan Italia Srl

Tecan Group Ltd

12 VETERINaRY TECAN JOURNAL 2/2012

The Fédération Nationale des Courses Françaises oversees the fight against doping in French horse racing through the Laboratoire des Courses Hippiques (LCH), based in the Paris suburb of Verrières-le-Buisson. The laboratory analyzes around 45,000 blood and urine samples annually, two thirds of which are from France, with the remainder from countries as far afield as Qatar, United Arab Emirates (UAE) and India, as well as from European racing authorities such as Holland, Belgium, Switzerland and Spain. In addition to routine drug screening, the laboratory provides counter analysis of ‘B’ bottles for other racing laboratories, is a reference laboratory for the Fédération Equestre International (FEI), and analyzes competition horse feeds for contaminants. The laboratory also analyzes a small number of human samples alongside the equine samples – about 500 a year – plus samples from dog

races and, more unusually, from camel races in the UAE and Qatar. This is backed by an active research program which helps to continually advance anti-doping control methods and keep LCH one step ahead of potential cheats.

During a race meeting, the race commissioner selects horses for drug screening. These include all race winners, as well as horses which may be exhibiting unusual behavior, underperforming or performing unusually well. The course vet verifies the identity of a horse from its microchip and the barcode on its passport, and records its sex, age and the discipline – trot or gallop – in which it has competed. Blood and urine samples – divided into two bottles, the ‘A’ and ‘B’ samples – are then assigned barcodes, sealed and dispatched to the laboratory. Patrick Cuvier, IT Manager at

LCH, described the sample reception process: “Urine samples arrive at the laboratory in 200 ml bottles and blood samples in 10 ml tubes. We scan the barcode and record the information provided by the vet, along with the number of vials per horse and the approximate sample volume. Each sample is then allocated a unique laboratory number to enable full sample tracking.”

LCH is constantly seeking to improve its analytical methods, and has recently invested in a Freedom EVO 200 workstation equipped with an eight-channel Liquid Handling (LiHa) Arm and custom -made carriers to enhance sample preparation for mass spectrometry (MS) analysis, complementing the laboratory’s existing Freedom EVO 75 system for PCR set-up and Freedom EVO 150 for ELISA processing. The new platform’s LiHa Arm – which uses Tecan’s recently released

The LCH team with the Freedom EVO 200

Horse racing is an immensely popular sport worldwide and, with huge sums of money riding on the results, maintaining the integrity of racing is crucial for jockeys, trainers, owners and racegoers alike, as is protecting the health of the sport’s valuable thoroughbred horses. In France, the Laboratoire des Courses Hippiques is taking advantage of Tecan’s Freedom EVO® platform to prepare samples for analysis by mass spectrometry in the battle to keep racing clean.

High stakes laboratory automation

13

5 ml disposable tips – enables efficient transfer of large sample volumes, and the laboratory can process up to 144 urines and 192 bloods a day in batches of 24 samples. Patrick commented: “Manually pipetting 45,000 samples per year is an enormous undertaking that is both tedious and time-consuming. Using the Freedom EVO allows us to decrease the time spent on repetitive pipetting, freeing staff to perform other tasks and enabling us to increase our sample throughput.”

Dr Ludovic Bailly-Chouriberry, Assistant Laboratory Manager, explained the process: “Samples are analyzed using one of seven different drug screens, each focusing on the detection of a particular type of compound. We use the Freedom EVO 200 to transfer sample aliquots into appropriate tubes and vials, which are then taken for analysis. Blood samples for ELISA are transferred directly to the Freedom EVO 150, which is equipped with four-channel LiHa and Robotic Manipulator

(RoMa) Arms, a HydroFlex™ washer, a four-slot MIO™ incubator/shaker and a Sunrise™ plate reader, enabling full automation of the assay and generating results in just a few hours.”

“One of the biggest challenges in this process is the difficult nature of urine and plasma samples,” continued Ludovic. “Blood samples must be allowed to settle naturally in a cold room before plasma can be extracted for analysis, and horse urine has a wide viscosity range. To cope with these issues, we needed a system that would provide accurate and precise pipetting while avoiding the risk of cross-contamination, as our analytical methods are extremely sensitive. The Freedom EVO’s liquid level detection function is an important aspect of this process security, enabling the system to quickly determine the amount of usable sample, which can vary from a few milliliters to 200 ml. The safe pathways feature in

Freedom EVOware® also helps to virtually eliminate the risk of cross- contamination by ensuring that the LiHa never travels over a sample or another piece of labware; it’s impressive and very convenient.”

And in the future? Ludovic imagines a system that has evolved to encompass the laboratory’s entire workflow, from sample reception and extraction through to analysis by mass spectrometry. “We could adapt our Freedom EVO platforms to provide sample preparation for all generations of MS systems, and would no longer need to keep sample preparation and analysis separate. It would really be a beautiful challenge,” concluded Ludovic.

To find out more on Tecan’s veterinary diagnostic solutions, visit www.tecan.com/veterinary

To find out more about LCH, visit www.fncf.fr

Sample bottles are loaded into custom-made carriers on the Freedom EVO

Aliquoting urine samples from 200 ml bottles using the LiHa Arm with 5 ml disposable tips

VETERINaRY TECAN JOURNAL 2/2012

14 REaDERS TECAN JOURNAL 2/2012

Developing new, efficient immunodiagnostic technologies

The Department of Biomolecular Engineering at the Kyoto Institute of Technology, Graduate School of Science and Technology in Kyoto, Japan, uses biochemical engineering to develop new technological applications that mimic biological functions. Dr Yoichi Kumada, Assistant Professor at the Department, is conducting research to develop new immunodiagnostic technologies based on scFvs immobilized on laboratory plasticware.

“Currently used immunodiagnostic technologies are mainly based on ELISA and bead assays, but there are technical limitations in the immobilization of the antibodies, so we are exploring ways to improve this,” explained Dr Kumada. “We focused on the polystyrene in plasticware that is commonly used for routine laboratory work, and identified a peptide tag that

recognizes the polystyrene substrate surface with a very strong affinity (polystyrene-affinity peptide: PS-tag). In addition, we decided to use scFvs; while normal antibodies are Y-shaped molecules consisting of a combination of four peptide chains, scFvs have just one chain. The merit of this is that they are relatively inexpensive to produce; conventional monoclonal antibodies are produced by animal cells, but recombinant antibodies that are generated by E. coli can be produced at a considerably reduced cost. Furthermore, they are relatively small, so they can potentially be used at a higher density, contributing to higher test sensitivity. Their molecular structure is also conducive to a more favorable alignment when immobilized. By gene recombination technologies, we designed an scFv with the PS-tag (PS-tagged scFv) and produced this using E. coli.”

In order to efficiently analyze and optimize the immobilization process, Dr Kumada chose Tecan’s Sunrise™ and Infinite® 200 microplate readers. He continued: “We use the readers to measure the density of the PS-tagged scFvs that have been immobilized on the substrate in 96-well microplates, and to evaluate antigen binding activity. To measure density, we stain the immobilized proteins and measure their absorbance while, to measure activity, we perform an ELISA-based reaction, then measure the resulting fluorescence. Using this data, we can calculate the relative activity per molecule of the PS-tagged scFvs, and evaluate whether the antibodies can be immobilized stably and at high density using our technology. Furthermore, to measure the availability or, in other words, the exposure of the antibody, we use a fluorescence-labeled antibody that detects our immobilized scFvs, and again measure the fluorescence. We have shown that PS-tagged scFvs can be immobilized on polystyrene plates at high density, highly favorably aligned and with high activity. Compared to the conventional whole antibodies (mAb), our PS-tagged scFvs can be manufactured at less than 10 % of the cost, and have improved test sensitivities by 10 to 100 times.”

Dr Kumada described the instruments’ strengths: “The Sunrise and Infinite give us more reliable readings compared to other instruments that we used previously, particularly giving good resolution even at low intensities. The Sunrise comes with Magellan™ software, which has a very useful absorbance kinetics function; our measurements are often taken at 30 second intervals for 30 minutes, and Magellan automatically plots the kinetics of each well on a chart. With the Infinite, we have found

Researchers at the Kyoto Institute of Technology chose Tecan’s microplate readers and microarray processing instruments to develop highly sensitive immunodiagnostic technologies based on immobilized single-chain variable fragments (scFvs).

Kyoto Institute of Technology

antibody PS-tag-fused scFv

Flexible linker

VH Chain

VL Chain

Antigen bindingFv fragment Antigen

Antigen

15REaDERS TECAN JOURNAL 2/2012

that the selection of the measurement area is very easy, and Tecan also provides excellent support for non-standard plates; we have only needed to make a call, and a script has been written to allow us to use multi-well plates of different formats.”

More recently, the Biomolecular Engineering laboratory has started using Tecan’s microarray-processing instruments to evaluate the antigen binding activity of the PS-tagged scFvs that have been immobilized on biochips. Dr Kumada continued: “We use Tecan’s HS400™ Pro hybridization station for automated processing of our scFv-immobilized biochips, including blocking,

washing, antigen-binding and fluorescence-labeled antibody binding. The same slide carrier used in the HS400 Pro is compatible with Tecan’s LS Reloaded™ scanner, so we can easily transfer the chips to the microarray scanner for fluorescence detection. We have found the scanner to have very good reproducibility, giving us confidence in our data from quantitative experiments, and its accompanying software makes it very easy to use.”

“Although DNA chips are often used these days, protein chips are somewhat behind in their development,” explained Dr Kumada. “A few are available for research, but products

for practical use are very limited, and I hope that the technologies that we are developing will help these products to become reality. We are continuing our research to develop immunoassays, biosensors and antibody arrays using PS-tagged antibodies, with the aim of implementing their practical applications.”

To find out more about Tecan’s microplate readers, visit www.tecan.com/detection

To find out more about the Kyoto Institute of Technology, visit www.kit.ac.jp/english

The Department of Biomolecular Engineering team at the Kyoto Institute of Technology. Front to back; Dr Yoichi Kumada, Dr Lee Joo-Young, Kagenari Yamakawa, Yusuke Emori

Antibody and the single chain variable fragment (scFv)

16 MICROaRRaYS TECAN JOURNAL 2/2012

Finding the right candidate

The complex nature of antibodies creates a number of challenges for the development of new biopharmaceuticals, as most antibodies will bind to multiple targets with varying affinity. Novartis’ Integrated Biologics Profiling group in Basel, Switzerland, is responsible for assessing the technical developability of candidate antibodies as potential biopharmaceuticals, and uses Protagen UNIchip protein microarrays to identify off-target effects.

Steffen Hartmann, Head of the Protein Developability Group, explained the process: “Our colleagues in research begin with a therapeutic target which they wish to direct an antibody against, then identify potential candidate antibodies for further investigation. We generally start with around 50 antibodies which show binding affinity for the target of interest, and profile each of these according to various criteria to assess the potential to further develop the

antibody for therapeutic use. An important aspect of this profiling is the identification of off-target effects, which are particularly important in drugs derived from biological macromolecules, as they usually bind to multiple targets. My colleague Jean-Marc Schlaeppi, Senior Investigator in the NIBR Biologics Center, became interested in protein microarrays as a potential method for quickly and conveniently identifying off-target binding activities, as this technique allows

Novartis is using an HS 4800™ Pro hybridization station to help profile and select antibodies for development into new biopharmaceuticals, relying on the system’s throughput and reproducibility to efficiently prepare Protagen UNIchip® microarrays for analysis.

Stefan Muellner and the Protagen team have extensive experience with Tecan equipment

17MICROaRRaYS TECAN JOURNAL 2/2012

a broad range of targets to be presented in a compact format and is suitable for multiplexing and automation.”

“Several life science providers offered protein microarrays at this time,” added Xavier Leber, the research associate responsible for the development and running of the protein microarray studies. “Jean-Marc began by assessing which of these best matched our needs, and the Protagen UNIchip performed very well in these initial tests. The Company’s staff were also very helpful – taking time to understand our exact requirements before making their recommendations – and so we were confident that the UNIchip was the best fit for our workflow. Although we piloted the use of the Protagen protein microarrays manually, the intention was to employ an automated hybridization system, and the experience that Protagen had already gained using Tecan’s HS Pro hybridization stations was also an important consideration.”

Stefan Muellner, CEO of Protagen, continued: “We have been using Tecan’s HS Pro hybridization stations in both our product development and contract research activities

since 2006. These were first introduced to eliminate the variability associated with manual assays, which is particularly important when developing a new commercial product, and vital to our role as a CRO. The innovative design of the HS Pro’s incubation chamber also provides very good mixing and distribution of reagents, improving the hybridization quality for all microarray types, and so we were confident that this system would be suitable for Novartis’ needs.”

Steffen Hartmann continued: “Because we wanted to get the hybridization set-up running as quickly as possible, we were keen to take advantage of the Protagen team’s experience and protocols with the HS Pro and, as Tecan has a very good reputation for high quality, reliable products, we were happy to purchase an HS 4800 Pro based on a recommendation from Protagen. We were then able to get up and running very quickly; Protagen provided the basic protocols, as well as a lot of technical information on running the UNIchip arrays, and the HS 4800 Pro’s user-friendly design made it easy for us to set up the instrument for our needs. Based on

The HS Pro allows the Novartis team to process up to 24 slides in parallel

A PowerScanner™ provides high throughput scanning of the UNIchip microarrays

our experiences, we also purchased a Tecan PowerScanner to perform high throughput scanning of the microarrays. This system perfectly complements our HS Pro, and was the only platform on the market which met our workflow requirements.”

Steffen concluded: “Our HS 4800 Pro has two extension units, allowing us to hybridize 24 slides in parallel, and our workflow has been designed to minimize hands-on time while offering excellent reproducibility and high quality results. This gives us the ability to quickly perform off-target binding studies for a broad range of candidate antibodies and to change the conditions as required, helping to minimize turnaround times and accelerate the overall candidate selection process.”

To find out more on Tecan’s HS 4800 Pro hybridization station, visit www.tecan.com/hs4800

To learn more about Novartis, go to www.novartis.com

To learn more about Protagen and the UNIchip protein microarrays, go to www.diagnostics.protagen.com

“The innovative design of the HS Pro’s incubation chamber improves the hybridization quality for all microarray types...”

18 SERVICE TECAN JOURNAL 2/2012

Caring for staff and patients; benefits all round with the FE500™ and a Te-Care™ Service Contract

Wentworth-Douglass Hospital in Dover, New Hampshire, USA – the Seacoast’s leading medical center – provides services to an area that also includes Eliot and South Berwick in Maine, and Lee and Barrington in New Hampshire. Darlene Bickford, Support Services Manager at Wentworth-Douglass, manages both the in-house and outreach phlebotomists, supporting them and the lab aids in the anatomic and clinical laboratories. The hospital’s in-house phlebotomists service all of the units on a three shift system, and they are also responsible for processing all specimens that come into the central laboratory, including those from a number of outreach laboratories, the busiest of which can have around 100 patients in a 12-hour time period.

Darlene explained: “As you can imagine, this is a very high throughput department and the main reason for buying the FE500 was to help us process the 1,600 or so samples that come from our outreach program and in-house patients each day. The system is installed in our main laboratory here at Wentworth-Douglass, centrally located between our support services and technical areas, and all our specimens – with the exception of stats which are sent straight through to the laboratory – are processed on the FE500.”

Darlene continued: “All the samples are barcoded, enabling the FE500 to communicate all information to our LIS.

The Department of Pathology and Laboratory Medicine at Wentworth-Douglass Hospital relies on an FE500 pre-analytical sample handling platform and a Te-Care Service Contract, ably supported by Tecan’s certified field service engineers, to ensure uninterrupted processing of blood samples for its clinical departments.

19SERVICE TECAN JOURNAL 2/2012

Clot detection is an important feature of the system, as is the fact that it also centrifuges samples. After labeling a secondary tube, the FE500 aliquots the specimen and places the tube into the appropriate carrier, where it is ready for collection by the technicians or phlebotomists. It’s a really slick, walkaway system. We don’t have to worry about forgetting samples that are in the centrifuge or have to wait around to unload them, we just put them on and walk away. We find the FE500 very easy to use and it copes easily with our high sample throughput. It has also reduced the risk of exposing our phlebotomists to blood-borne pathogens through splashing, and improved our turnaround times.”

For such an important high throughput, 24/7 application, maximum up-time is essential and covered, in this case, by a Te-Care Service Contract. Darlene added: “Our service contract includes preventive maintenance and, on the occasions when we have needed additional support, I have simply sent an email and had an immediate response. Often, they can talk us through any issues and solve problems over the phone but, if a site visit is necessary, an engineer always arrives very quickly. The response from Tecan is tremendous.”

“In the beginning it was a big transition from manual processing to using the FE500 and, at this time especially, Tecan’s support was excellent, providing us with training and

helping us through the implementation. Bringing in this new analyzer for the front end was a huge change but was a really good move. Processing everything manually would be very time-consuming, taking our phlebotomists away from caring for patients, and we just can’t imagine being without the FE500.”

To find out more on Tecan’s Te-Care Service Contracts, visit www.tecan.com/services

To find out more about Wentworth-Douglass, visit www.wdhospital.com

The Wentworth-Douglass team with the FE500. Front row, left to right: Bob Rickard, Dottie Robichaud, Karen Martin and Tammy Fermino; back row, left to right: Arielle Vachon, Darlene Bickford, Patty Sheckler and Katie Gosselin

Wentworth-Douglass Hospital

20 REaDERS TECAN JOURNAL 2/2012

A real winner for celiac patients

Celiac disease is the result of intolerance to gliadin, a constituent of the cereal protein gluten, requiring sufferers to strictly adhere to a lifetime gluten-free diet. A consequence of this is that the level of gliadin in gluten-free foods must be carefully controlled to ensure the food safety for celiac patients, making an easy and reliable method of gliadin analysis essential for food manufacturers producing gluten-free foodstuffs. To prevent celiac patients suffering adverse reactions to food, gluten is included in regulations relating to food labeling, with EC legislation stipulating that gluten-free products must not exceed a level of 20 mg/kg of food. Manufacturers must check for gluten contamination throughout the food production process and a key part of this is rapid testing of incoming raw materials.

Researchers in the Sensors and Biosensors Group at the Universitat Autònoma de Barcelona, Spain, have taken advantage of Tecan’s Sunrise™ absorbance reader and Magellan™ V4.0 software to establish a novel magneto-ELISA for gliadin, based on optical detection. Maria Isabel explained: “There is an increasing demand for rapid, simple and low cost techniques for accurate food analysis in decentralized analytical situations. As a result, there is a great deal of interest in the development of biosensors for these applications, due to the high sensitivity and excellent specificity they offer. Immunosensors, in particular, are proving promising alternatives to the existing immunochemical tests that ensure food safety from the farm to the table.”

“Recent advances have enabled magnetic beads to be used as a support for immunoassays. This significantly improves the performance of the immunological reaction, as the surface area is increased and the beads can be easily manipulated using a magnetic field. Assay kinetics are more rapid because the beads are in suspension, and the improved washing and separation steps help to minimize matrix effects in complex samples. In addition, the magnetic beads can easily be integrated into microfluidic devices and cartridges. We have taken advantage of this to develop an electrochemical magneto immunosensor for the sensitive detection of gliadin – and small gliadin fragments – in natural or pretreated foods such as skimmed milk and gluten-free beer. The immunological reaction is performed on magnetic beads

The Tecan Award 2011 built on last year’s success and saw our customers submit a plethora of innovative applications for our detection equipment. Competition was fierce, with many ingenious entries, and third place was eventually awarded to Maria Isabel Pividori from the Universitat Autònoma de Barcelona, for her work on the detection of gliadin, a constituent of the cereal protein gluten.

The Biosensors and Bioanalysis team. Left to right; Susana Liébana, Tamara Laube, Delfina Brandao, Soledad Carinelli, Daniela Santos, Rey Bundalian, Isabel Pividori

Third place

21REaDERS TECAN JOURNAL 2/2012

as a solid support, enabling gliadin to be successfully immobilized and oriented onto the magnetic beads. The biorecognition strategy is based on a competitive assay – using commercially available antibodies labeled with horseradish peroxidase (HRP) – and the modified magnetic beads are captured onto the surface of a magneto electrode based on graphite-epoxy composite (m-GEC) for electrochemical detection. This low cost biosensor allows gliadin analysis to be performed easily throughout the food chain, enabling control of each stage in the production process.”

Maria Isabel continued: “To assess the performance of this assay, we compared it with a novel magneto-ELISA, using optical detection performed on the Sunrise plate reader. As with the immunosensor, we immobilized gliadin on magnetic beads and used HRP as the label for the enzymatic reaction, optimizing both direct and indirect competitive assays. The ELISAs were performed in 96-well microplates, using a magnetic separation plate to isolate the supernatant before measuring the absorbance in the Sunrise reader. The recovery values using both methods were very good; the limit of detection for the magneto-ELISA was excellent and, as with the magneto immunosensor, we were able

todetectlevelsof20µg/kgoffood,whichis 1,000 times lower than the EC-specified 20 mg/kg limit of detection for gluten-free food.”

“I particularly like the versatility of the Sunrise reader,” concluded Maria Isabel. “It enables magneto-immunoassays to be performed in a variety of different formats for multiple applications – such as evaluating protein coupling to magnetic beads and nanoparticles – as well as assessment of different transducer materials for biosensing purposes. It also allows us to quickly and easily optimize reagents and assay parameters, making it ideal for research applications.”

Schematic representation of experimental procedures for A) magneto-ELISA and B) electrochemical magneto immunosensor

Tamara Laube inserts a plate into the Sunrise reader

Full details of this study can be found in: Laube T et al. Biosens Bioelectron, 2011, 27, 46-52.

To learn more about Tecan’s Sunrise absorbance reader, visit www.tecan.com/sunrise

To find out more about Maria Isabel Pividori at the Sensors and Biosensors Group, visit webs2002.uab.es/ipividori/

“This low cost biosensor allows gliadin analysis to be performed easily throughout the food chain, enabling control of each stage in the production process.”

Experimental procedures

(B) Electrochemical magneto immunosensor

amperometric unit

(A) Magneto-ELISA

mag

net

m-GEC electrode

Abs (λ=450nm)

magnet

22 CELL bIOLOgY TECAN JOURNAL 2/2012

Capturing the ‘dynamics’ of automated cell culturing

CDI, based in Wisconsin, USA, is currently the world’s most innovative stem cell company, as voted by The Wall Street Journal in its 2011 Technology Innovation Awards. Founded to commercialize ground-breaking technology developed by stem cell pioneer James A. Thomson, CDI produces industrial quantities of differentiated human cells for basic research, drug discovery and development, and potential therapies to advance personalized medicine. To date, CDI has had great success launching iCell® Cardiomyocytes, iCell Endothelial Cells and iCell Neurons, with hepatocytes and many other cell prototypes already in development. The Company will also soon be launching its MyCell™ Custom Services for custom reprogramming.

Scaling up the production of pluripotent stem cells and directing differentiation to

different cell types is technically challenging, labor-intensive and has inherent process variability. From an economic, regulatory and process control perspective, manual processing of stem cells is not a viable long-term solution. With these challenges in mind, researchers at CDI began to work with automated systems in order to standardize the maintenance and expansion of hiPSCs, and the subsequent differentiation into HPCs. To improve technical limitations, mainly the lack of throughput, a more sophisticated liquid handling system, Tecan’s Cellerity, was purchased for the task.

The system, a Cellerity500, is based on a Freedom EVO® 200 liquid handling platform, and includes a LiCONiC™ STX 500 automated incubator with a capacity for 500 plates, a media storage fridge, an AutoLoader™ for loading flasks, a Cedex™ cell counter (Innovatis, Roche), spinner flasks for expansion and seeding of suspension cells, a Robotic Manipulator Arm to handle plates and an eight-channel fixed tip Liquid Handling Arm. The Cellerity also includes a positively pressured HEPA filtered environment in a BSL2-compliant class

100 sterile room for operations under GMP guidelines. Cells are maintained under sterile conditions in automation-friendly Corning® RoboFlasks®, which have an area of slightly less than 100 cm2 and have a pierced septum for automated access. The system is disinfected using a sporicidal solution at the end of each run to prevent cross-contamination between cell lines.

Nick Seay, Chief Technology Officer at CDI, explained: “We are using the unique capabilities of the Cellerity robotic cell culture system to successfully generate blood precursor cells from hiPSCs that are often found only in sparse numbers in post-natal tissues. This robust and efficient automation system delivers consistency and other systematic improvements to our processes, allowing us to optimize differentiation to produce multipotent (CD43+/CD34+) HPCs and other defined cell types from blood.”

The automated splitting of hiPSCs was made possible by the use of ROCK inhibitors and its effect on improving the cell viability of individualized cells. Cells can be lifted and individualized with trypsin, and plated

Researchers at the innovative stem cell company Cellular Dynamics International (CDI) have developed a novel, serum-free directed differentiation protocol, based on Tecan’s Cellerity™ automated cell culturing system. The new method enables the maintenance of a uniform starting population of human induced pluripotent stem cells (hiPSCs), leading to the generation of hematopoietic precursor cells (HPCs).

Research team members: Nick Seay, Deepika Rajesh, Giorgia Salvagiotto, Elisabeth Enghofer, Christie Munn, Candice Lindstrum and Sarah Burton

23CELL bIOLOgY TECAN JOURNAL 2/2012

without centrifugation as single cell cultures in the presence of trypsin inhibitor in mTeSR media. The cells are then expanded under feeder-free conditions on matrigel-coated RoboFlasks. This technique, developed at CDI (patent pending), eliminates the need for mechanically detaching the cells from the surface of the plates, and the system is capable of generating 5-6 billion hiPSCs in a weekly process.

Critically for the process, automation does not alter the genetic stability of these cells, nor their potential to form blood precursor cells. The serum-free differentiation is achieved on matrix proteins in the presence of cytokines. Aliquots of a heterogeneous HPC population are placed in various defined cytokine cocktails for further selective differentiation to a myeloid (generating macrophages and dendritic cells), megakaryocyte (platelets), or erythroid (red blood cells) lineage following an additional two to three weeks in culture.

Nick added: “We have also used the Cellerity to screen over 2,000 small molecules, identifying those that boost the generation of blood-forming precursors from stem cells and improving the efficiency and quality of the blood precursor stem cells even further. Overall, the project has significantly enhanced our scientific understanding, enabling us to control the growth, maintenance and differentiation of stem cells into blood precursor cells, as well as other specialized cell types, with increased yield and precision. The ability to expand stem cells and produce large numbers of differentiated cell types by automation is a real hallmark of our success in this developing field.”

To find out more on Tecan’s cell biology solutions, visit www.tecan.com/cellbiology

To learn more about CDI, go to www.cellulardynamics.com

Cells maintained in a RoboFlask

Aggregates generated in a 96-well plate

Differentiation for 12 days

Addition of small molecules (All steps performed on the Cellerity)

High throughput system overview

Hematopoietic precursor cell

The Dittmer Lab is part of the Department of Microbiology and Immunology and Lineberger Comprehensive Cancer Center at the University of North Carolina (UNC), and focuses on the investigation of viral cancers, such as those caused by Kaposi’s sarcoma-associated herpesvirus (KSHV). KSHV is responsible for one of the most common AIDS-associated cancers, posing a significant risk to HIV-positive or immuno-compromised individuals. The Group uses qPCR profiling to study various aspects of KSHV, as postdoctoral research associate Pauline Chugh explained: “We use three main strategies to investigate viral tumorigenesis – viral gene expression, microRNA (miRNA) expression and viral load testing of clinical samples – based on 96 primer qPCR arrays. Performing the assays manually would be very labor-intensive, and so we use automation to provide the throughput necessary for these studies.”

The Dittmer Lab relies on a Freedom EVO 150 workstation equipped with an eight-channel Liquid Handling (LiHa) Arm to ensure accurate and reliable assay plate set-up in a 384-well format. This platform is linked to a Roche LightCycler 480 Real-Time PCR System (LC480) to provide walkaway automation of qualitative and quantitative nucleic acid detection by real-time PCR. Pauline continued: “This set-up gives us the ability to perform automated analysis of up to 750 gene sequences per sample in just one day, allowing us to, for example, look at the whole miRNA library following infection with the virus. Before purchasing the Freedom EVO platform we used a basic, single channel pipetting robot, which was not capable of reliable automation on the scale required for this kind of investigation, and required manual transfer

of plates to the LC480. In contrast, we can now load 60 individual arrays – four arrays per 384-well plate – onto the Freedom EVO and leave the system running overnight. It’s basically generating data while you sleep.”

“Having the ability to multitask during working hours has also significantly improved our productivity, allowing us to perform other experiments or analyze data while the instrument is running, and having confidence in the platform’s pipetting accuracy and process security is vital to this. We have run numerous quality control tests to make sure that the results the system produces are robust, and have seen no systematic errors or issues, obtaining good standard deviations for the data.”

“The system’s accuracy and reproducibility have also allowed us to miniaturize many of our protocols, which offers significant cost savings in terms of reagents. For example, we have been able to reduce the total volume for our miRNA assays from the manufacturer-recommended20µltojust 6 µl.Optimizingtheliquidclasseshasallowed us to significantly reduce the amount of very expensive miRNA primer

Dr Pauline Chugh

The Dittmer Lab at the University of North Carolina is investigating the biology of viral cancers using a Freedom EVO® 150 with an integrated LightCycler® 480 Real-Time PCR System. This versatile set-up provides automated qPCR -based analysis for viral gene and microRNA expression profiling, as well as viral load testing of clinical trial samples.

A powerful combination for qPCR

24 gENOMICS TECAN JOURNAL 2/2012

University of North Carolina

“The system’s accuracy and reproducibility have allowed us to miniaturize many of our protocols, which offers significant cost savings in terms of reagents.”

25gENOMICS TECAN JOURNAL 2/2012

The Roche LC480 is linked directly to the Freedom EVO workstation

required per assay – effectively doubling the number of assays we can perform – with no loss of accuracy, and Tecan’s engineers have been instrumental in realizing these benefits. As most of our experiments use very similar methods, Tecan worked with us to create a script for one of our more complex protocols when the platform was first installed, and spent several days training us to adapt and optimize it for our other assays.”

To minimize the risk of contamination, miRNA primers are stored in Matrix SepraSeal® tubes with pre-slit, self-sealing septum caps which can be pierced by the

LiHa Arm’s fixed tips. Pauline commented: “Although the need to wash the pipette tips thoroughly after every dispense makes the protocol longer than our methods which do not use Matrix tubes, the enhanced process security and cost savings from using lower primer volumes more than offset this, and we are still able to generate data far quicker than it can be analyzed.”

The flexibility and performance of the Freedom EVO has also led to the Dittmer Lab’s recent purchase of a second instrument equipped with a MultiChannel Arm™ 96, for use on other projects. “This platform

is intended for multiple applications, and allows us to take advantage of the benefits of automation without disrupting our ongoing work. We are also inviting members of other groups and departments to use this flexible system, encouraging new collaborations and furthering our research,” Pauline concluded.

To find out more on Tecan’s genomics solutions, visit www.tecan.com/genomics

To find out more about the Dittmer Lab, visit www.med.unc.edu/microimm/faculty/dittmerlab

26 MICROaRRaYS TECAN JOURNAL 2/2012

Automation aids peptide biomarker discovery

JPT Peptide Technologies in Berlin, Germany, has developed a proprietary peptide microarray technology – PepStar – based on high throughput synthesis and immobilization of peptides onto glass slides. The Company uses these peptide arrays and Tecan’s HS 4800 hybridization station to profile humoral immune responses in patient sera samples. Dr Holger Wenschuh, CEO of JPT, explained: “These profiles help in the search for diagnostic biomarkers of infections, autoimmune diseases, cancers and allergies. We receive samples from studies run by our collaborators in biotechnology companies, the pharmaceutical industry and the medical profession, searching for new peptide biomarkers which are indicative of specific stages of disease, which can be used as tools to monitor the success of a therapeutic intervention or to establish an immunological rationale for clinical findings.”

Holger continued: “For each study, we optimize the assay conditions before we

begin testing hundreds of samples in parallel; we have to be able to prove that our protocols are reproducible and reliable. The need for profiling clinical samples under robust experimental conditions was the primary reason for automation – although throughput was also an issue – as manual hybridization introduces variances, even when standard operating procedures are followed exactly. These are significantly reduced by automated procedures and therefore, for the past three years, most of our protocols have been run on the Tecan hybridization station. After evaluating several systems, with particular regard to handling procedures and reproducibility, we concluded that the HS 4800 hybridization station was the best instrument for our purposes. Tecan was also very willing to work with us to establish instrument protocols specific for our needs.”

The initial step in peptide microarray manufacture is the use of bioinformatics to generate complex, specifically tailored

Researchers at JPT Peptide Technologies are using Tecan’s HS 4800™ hybridization station to improve throughput and give a high level of reproducibility for patient sera screening with the Company’s own PepStar™ peptide microarray technology.

Holger Wenschuh, CEO of JPT

Janina Seznec using JPT’s hybridization station

“For each study, we optimize the assay conditions before we begin testing hundreds of samples in parallel; we have to be able to prove that our protocols are reproducible and reliable.”

27MICROaRRaYS TECAN JOURNAL 2/2012

peptide libraries that result from the human proteome or the proteomes of pathogens; these libraries typically reflect antigens related to diseases of interest. Chemical synthesis of these peptides using high throughput technology produces tens of thousands of peptides in microplates, which undergo rigorous quality control before being immobilized onto the slide surface to produce peptide microarrays. Critically, this immobilization is site-specific, yielding purified peptides with the same orientation on the slide.

The peptide microarrays are placed in the hybridization station and incubated with appropriately diluted patient samples, allowing circulating antibodies in the samples to bind to immobilized peptides with the correct linear target epitope. Any unbound antibodies are removed by a series of washes before addition of a fluorescently-labeled reporter antibody, which binds to any captured antibodies from the patient’s

serum. Holger continued: “Applying peptide microarray technology in large studies – using hundreds of samples to broadly profile humoral immune responses – is quite a new area, but we were able to set up the Tecan hybridization station for this application pretty quickly, thanks to its user-friendly interface. Establishing automated protocols and standard operating procedures – either through creating new protocols or transferring existing manual ones – was achieved within two months, allowing us to quickly bring the system into routine operation. The cohort sizes vary, but a typical batch of 100 to 200 patient samples will take about two weeks to process using this set-up.”

“Although most of our work focuses on the discovery of new markers and serological profiling of clinical samples, the PepStar platform is now being used in enzymatic profiling with cell lysates and a variety of other samples, which also takes advantage

of the hybridization station’s reproducibility. This technique is helping us to identify appropriate substrates for enzymes with a role in pathogenesis, allowing development of high throughput screening assays for enzymatic activities indicative of diseases, a technique which is becoming increasingly useful in the field of therapeutic drug discovery.”

Holger concluded: “We have been very impressed by the performance of our hybridization station, and – as a leader in the peptide microarray field – have recommended the HS Pro to several of our customers and collaborators, who are now using these instruments very successfully in their laboratories.”

To find out more about Tecan’s microarray solutions, visit www.tecan.com/microarray

To learn more about JPT Peptide Technologies, visit www.jpt.com

The team at JPT

28 DRUg DISCOVERY TECAN JOURNAL 2/2012

The right tools to drive drug discovery

As the name suggests, the DMPK Group in the Vanderbilt University Medical Center Department of Pharmacology’s Center for Neuroscience Drug Discovery focuses primarily on targets aimed at treating neurological disorders, such as Parkinson’s Disease, Alzheimer’s Disease, schizophrenia and various diseases of cognitive impairment, as well as Fragile X Syndrome, a leading genetic cause of autism, and anxiety disorders. The Group has a special interest in modulating metabotropic glutamate receptors and muscarinic receptors to potentially treat these neurodegenerative and neurocircuitry conditions with greater therapeutic benefit and fewer side effects than current therapies. For this, and other similar projects, it partners with several companies and organizations also working in these areas and is heavily involved with the National Institute of Mental Health and NIH’s molecular libraries program (MLPCN).

In 2010, with the transition to three lead optimization programs, the workload of the DMPK Group was approaching a critical level and inevitably errors in manual handling and throughput were an issue. In order to tackle the high throughput nature of some of the assays, plans were put in place to build a contemporary drug metabolism facility. The laboratory was already equipped with some automated pipetting systems, but these were not capable of achieving the necessary throughputs and the experimental integrity – consistent temperatures and consistent shaking – so the hunt for alternatives began. A key consideration was the types of in vitro systems routinely used – microsomes and liver S9 fractions – which are of such a consistency that they need to be shaken at just the right RPM to prevent settling and precipitation.

Dr J. Scott Daniels, Assistant Professor of Pharmacology and Director of DMPK for the VCNDD, explained why the Group decided on the Tecan Freedom EVO 200 early in 2011: “Firstly, the instrument is a quality product that is easy to operate and it was competitively priced, which has to be an issue. However, we were also very impressed with how keen Tecan was to work with our laboratory at the ground level, to help us build up our HT DMPK operation. Although many suppliers might promise a high level of customer service, our experiences with Tecan support have so far been exemplary and started before the instrument was even in place. Once everything was ordered, Tecan’s first step was to provide our laboratory manager, Ryan Morrison, with software to start writing the programs, with several hours of software support with an application specialist. Ryan was able to build preliminary workflows so we could hit the ground running when the instrument was installed and had it running online very quickly.”

Ryan described the system the Group now runs on a daily basis: “We have a Freedom EVO 200 workstation fitted with three robotic arms – an eight tip Liquid Handling Arm, a MultiChannel Arm™ 96 and a RoMa Arm. On the deck, there is an incubated six-position IKA® shaker that Tecan had customized for us so that it has a really tight radius, plus a Safire2™ microplate reader for a high throughput kinetic solubility assay. There is also an additional incubator, a linear shaker incubator that sits on the back of the deck, and hotel stacks for planned Caco-2 transporter assays. For ease of use, we are using all disposable tips. In the initial stages, I was very impressed with the 3D modeling

A Freedom EVO® 200 dedicated to ADME screening has transformed the workflow of the Drug Metabolism Pharmacokinetic (DMPK) Group at the Vanderbilt Center for Neuroscience Drug Discovery (VCNDD), creating a high throughput contemporary ADME facility that is driving drug discovery forward.

Scott Daniels, Director of the DMPK Group

29DRUg DISCOVERY TECAN JOURNAL 2/2012

software and the 3D renderings of how the workstation would look with all the modules that we would need to run our assays to plan out the space on the deck. Sitting down with the Tecan application specialist and having the time before installation to get used to the programming was really helpful.”

“We currently have three high throughput in vitro ADME screening assays running on the system: an intrinsic clearance assay, a plasma protein binding assay with RED (rapid equilibrium dialysis) plates, and a cocktail-style cytochrome P450 IC50 assay. We are looking at adding two more assays in the near future and, although the assays are not running concurrently at present, we could very easily run them on the same day if our throughput increased, as laid out on the 3D simulator in the development stages.”

Scott continued: “In just one day, our Tecan system can help us screen compounds for their metabolic stability, their extent of protein binding, and whether there is the potential for drug-drug interactions. It has revolutionized our throughput in in vitro subcellular work, and we’re now looking at its potential in screening at a molecular level.”

“Overall, our system is very flexible, very efficient and has tightened up our reproducibility and error rate, saving considerably on compounds, microsomes and FTE time, simply because we no longer need to repeat failed Tier 1 assays. Another big advantage is that it builds in capacity, freeing up time for other bench work, data crunching or the next step of detailed PK Tier 2 assays.”

He concluded: “From struggling with single pipettors on the bench, we have progressed to a fully automated linear platform with which we can perform several bread and butter assays every single week to help drive the drug discovery pipeline. That is what we wanted to achieve, to drive the science, and to take risks in novel targets in a way that companies just cannot do given the current economic climate. We had a mission and we tackled it, and Tecan really lived up to its promise, partnering with us to make our facility into an efficient, high throughput contemporary ADME group.”

To find out more on Tecan’s options for ADME screening solutions, visit www.tecan.com/adme

To find out more about VCNDD, visit www.vcndd.com

3D modeling helps Ryan Morrison to maximize use of the available deck space

The laboratory’s workflow is centered around the Freedom EVO’s high throughput capabilities

Tammy Santomango shows the Group’s Freedom EVO, equipped with disposable tips for maximum sample security

Tecan Freedom EVO 200 workflow

“In just one day, our Tecan system can help us screen compounds for their metabolic stability, their extent of protein binding, and whether there is the potential for drug-drug interactions.”

Inform Medicinal Chemistry for

DMPK optimization

Series of compounds from

Medicinal Chemistry

Optimized compounds for advancement to

proof-of-concept studies

DMPK Screening on Freedom EVO 200

Liver metabolic stability

Plasma protein binding

Drug-drug interaction

(cocktail P450)

A fresh approach to clean water

Researchers in the Institute of Applied Microbiology at Giessen University, Germany, have developed a microplate-based assay for online testing of the effects of metal nanoparticles – including silver, iron and palladium – on bacteria and organic micropollutants. Michael Bunge, senior scientist in the Nanobioengineering group, explained: “It is clear that some metal nanoparticles are toxic to a variety of

microorganisms, but we don’t know how and to what extent they affect microbial growth and survival, and there are no standard methodologies to assess this accurately. We have, therefore, developed a straightforward technique that simultaneously cultivates microbes and uses the Infinite M200 microplate reader to monitor the effects of different sizes and compositions of nanoparticles on bacteria, specifically

enterobacteria and strains that exhibit antibiotic multi-resistance.”

“We began by measuring at specific time and end points, but soon discovered that we needed to look at the whole dynamic response of bacterial cultures. For example, some bacteria exhibit quite extended lag phases and didn’t even start to grow until 36-48 hours, while others have a very erratic pattern of growth. These observations may be highly significant, yet are undetectable using the more traditional microbiological end-point criteria, such as counting colony forming units or measuring inhibition zones, making continuous measurements vital.”

Michael continued: “We chose an Infinite M200 for this application because it is a monochromator-based reader, making it easy to adjust the wavelength down to a resolution of 1 nm – which is particularly important for nanoparticle suspensions with a high background signal – and I knew from previous experience that Tecan systems were reliable. The system itself is very easy to use, and it is saving us so much time on repetitive, laborious tasks. We also have the option to combine it with fluorescence techniques to assess the viability of cells in real time, or to add an injector module for automated liquid handling, should the need arise.”

The Nanobioengineering group (left to right): Karsten Theophel, Santosh K. Sandhi, Michael Bunge, Lara Neumann, Veronika Schacht, Victor Cheunuie-Ambe and Nassim Sahragard

Microbiologists at Giessen University are using an Infinite® M200 microplate reader to look at the potential of engineered metal nanoparticles for treating water, inhibiting the growth of bacteria in drinking water at the point of use, and eradicating pharmaceuticals and other micropollutants from waste water.

30 REaDERS TECAN JOURNAL 2/2012

Second place

31TaLK TO TECaN TECAN JOURNAL 2/2012

Leading the debateRemote support is a controversial subject, for service providers as well as customers, but Tecan’s US helpdesk has taken a long hard look at the benefits it brings to all parties and, in the last year, has increased the fix rate by telephone from 60 % to 71 %.

Remote support is not a simple solution; without a really skilled helpdesk and technical team it’s hard to achieve. At Tecan US, we have a mix of nine skilled individuals based in North Carolina, from computer specialists to laboratory scientists from all disciplines, on call 24 hours a day, 365 days of the year. Each one has undergone an intense 18-month learning phase in preparation for any call that might come their way and, if they can’t help straightaway, there will certainly be a colleague close at hand who can.

In 2011, with the help of this knowledgeable team, we decreased our field despatches by over 1,000 service calls, getting our customers up and running much faster than would previously have been possible, and saving considerably in costs for both the customer and Tecan. Hardware problems might still need a visiting engineer, but even diagnosing an issue over the phone can save time and help in preparation. When it comes to application or software issues, we can often solve things quickly by phone without the customer even needing to wait for a visit. With mutual trust and understanding, many customers have come to recognize that giving Tecan remote access to their instruments on these occasions, even if that means through firewalls, can give them instantaneous diagnosis and, in many cases, a quick fix there and then. This is truly a win-win situation and the way forward for the Tecan service organization.

Do you have experience of the advantages remote support can give? Send your comments to talk@tecan.com

Craig Williamson, VP Service and Consumables Tecan US, Inc.

“So far, we have applied this method to drinking water and we are working closely with companies interested in applying water filters directly to taps and other drinking water supplies in remote regions, especially in less developed areas of the world. Another potential application we would like to explore is the purification of waste water, particularly from hospitals, which contains many pharmaceuticals that are difficult to eradicate from water supplies, despite the efforts of treatment plants. We are hopeful that metal nanoparticles can be used to significantly reduce the recirculation of these and other micropollutants.”

To find out more on Tecan’s monochromator-based Infinite M200 microplate reader, visit www.tecan.com/infinite200

To find out more about the Institute of Applied Microbiology, Giessen University, visit www.uni-giessen.de/cms/fbz/fb09/institute/mikrobiologie?language_sync=1

Headquarters: Tecan group Ltd., Seestrasse 103, CH-8708 Männedorf, Switzerland T +41 44 922 88 88 F +41 44 922 88 89 info@tecan.com

Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276Design: OTM/London www.otmcreate.comPhotography: Marc Wetli/Zürich www.wetli.com, Günter Bolzern/Zürich www.bolzern.net, Susanne Völlm/Zürich www.susannevoellm.chEditor in Chief: Tecan Trading AG, Cornelia KegeleProject Lead: Tecan Trading AG, Cornelia Kegele/ Tanja Sager/Antonietta AlloccaEditor: kdm/UK www.kdm-communications.comPrint: DAZ Druckerei Albisrieden AG/Zurich www.daz.chAddress: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, journal@tecan.com, www.tecan.comTo register for the Tecan Journal please go to www.tecan.com/journal© 2012 Tecan Trading AG, Switzerland, all rights reserved.

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

www.tecan.com

Tecan are pioneers in automated liquid handling and innovative life science solutions. For over 30 years we continue to enable and support our customers to make the world a healthier and safer place.

32 EVENTS 2012 TECAN JOURNAL 2/2012

americas

Canadian Society of Forensic Science 58th Conference Burnaby, BC, Canada 08 – 11 May 2012ISBER International Society for Biological and Environmental Repositories 2012 Annual Meeting & Exhibits Vancouver, BC, Canada 15 – 18 May 2012

2012 National Medicinal Chemistry Symposium Oro Valley, AZ, USA 20 – 23 May 2012

60th ASMS Conference on Mass Spectrometry & Allied Topics Vancouver, BC, Canada 20 – 24 May 2012

AACC Annual Meeting and Clinical Lab Expo Los Angeles, CA, USA 15 – 19 July 2012

Cancer Cytogenomics Microarray Consortium Conference Chicago, IL, USA 01 Aug 2012

asia and Pacific

4th Annual World Pharmaceutical Summit 2012 Shanghai, China 06 – 08 June 2012

2012 SLAS Asia Conference and Exhibition Shanghai, China 19 – 21 June 2012

MEDIX Medical Device Development & Manufacturing EXPO Tokyo, Japan 20 – 22 June 2012

Analytical China 2012 Shanghai, China 16 – 18 Oct 2012

Europe, Middle East and africa

ESHG European Human Genetics Conference 2012 Nürnberg, Germany 23 – 26 June 2012

EAFS European Academy of Forensic Science 2012 Den Haag, Netherlands 20 – 24 Aug 2012

ELRIG European Laboratory Robotics Interest Group Drug Discovery Manchester, UK 05 – 06 Sept 2012

Meet Tecan at these events

Tecanaustria +43 62 46 89 33 belgium +32 15 42 13 19 China +86 21 220 63 206 Denmark +45 70 23 44 50 France +33 4 72 76 04 80 germany +49 79 51 94 170 Italy +39 02 92 44 790 Japan +81 44 556 73 11 Netherlands +31 18 34 48 17 4 Singapore +65 644 41 886 Spain +34 93 490 01 74 Sweden +46 31 75 44 000 Switzerland +41 44 922 89 22 UK +44 118 9300 300 USa +1 919 361 5200 Other countries +43 62 46 89 33