Tecan Journal Edition 01/2012

Tecan Journal Life Sciences and Partnering Business

Edition 1/2012

Braving the elements in Norway with Te-Care™ A complete service package for better patient care page 17

Reinventing drug titration page 6

Making sweet music: art and science join forces for the LabNotes project pages 18-19

Taking special care of dangerous pathogens pages 26-27

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CEO WELCOME TECAN JOURNAL 1/2012

Welcome to 2012

Welcome

We are already looking ahead to another busy and promising year here at Tecan, with the unveiling of some exciting new products at this year’s SLAS Conference in San Diego, California. This event gives the international scientific community its first chance to see technological breakthroughs that will help to push the boundaries of laboratory automation, and this year will be no exception with a number of new product offerings including the introduction of Tecan’s air displacement pipetting solution for the Freedom EVO® platform. This revolutionary technology will complement our liquid displacement technology, allowing Tecan to offer a choice of pipetting systems to best suit the needs of your applications.

Details of many of these new products and technologies can be found in this issue of the Tecan Journal – including the new Infinite® M1000 PRO with AlphaScreen® technology – as well as the usual mix of news, events and customer testimonials describing how Tecan systems best solve the needs of our customers.

We also have some of the highlights from the 2011 Tecan Symposium held in Shanghai, China.

We hope you enjoy the issue, and look forward to a prosperous and successful 2012 together.

Thomas Bachmann, CEO

CONTENTS TECAN JOURNAL 1/2012

page 6

Reinventing drug titration

2

CEO Welcome

4

ecan to launch Air LiHa for T Freedom EVO platform

pages 20-21

4 5 ml disposable tips now available for LiHa Arm 5 Tecan launches Infinite M1000 PRO with AlphaScreen and AlphaLISA® technology 6 Reinventing drug titration 7

ustomers benefit as Tecan C Australia becomes a subsidiary

8-10 4th Tecan Symposium enjoys Chinese success 11

hina embraces a whole new C dimension in blood safety

12-13 Successful isolation and expansion of human primary cells

pages 14-16

Biobanking clinical cytology samples in Sweden

Contents

pages 26-27

14-16 Biobanking clinical cytology samples in Sweden 17 Braving the elements in Norway with Te-Care™: a complete service package for better patient care 18-19 Making sweet music: art and science join forces for the LabNotes project 20-21 Unwinding the genetic clock 22-23 A new dimension in plate preparation for 3D cell assays 24-25 High throughput hybridoma generation 26-27 Taking special care of dangerous pathogens 27 Leading the debate 28 Events

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PRODUCT NEWS TECAN JOURNAL 1/2012

Tecan to launch Air LiHa for Freedom EVO® platform With over 30 years of experience in advanced liquid handling for life sciences, Tecan understands that flexibility is crucial to making laboratory automation work for you.

Our Flow­Thru™ Technology-based liquid displacement systems and automation expertise have established Tecan as a market leader in laboratory liquid handling solutions,

and we will soon be offering even greater choice by expanding our portfolio to include an air displacement pipetting arm for our Freedom EVO workstations.

Air

Liquid

Air and liquid displacement pipetting systems each have their own advantages.

Liquid displacement offers a choice of fixed or disposable tips, a large dynamic range and ultimate system cleanliness, whereas air displacement eliminates the need for system liquids and associated daily maintenance operations. The new Air Liquid Handling Arm™ (Air LiHa) for the Freedom EVO platform will provide our customers with greater versatility than ever before. By uniquely offering a choice of pipetting technologies on the same platform, or even combining both air and liquid displacement on a single workstation, Tecan will be able to provide unrivalled flexibility to suit the needs of your application and laboratory workflow and increase productivity.

The new Air LiHa will provide a choice of pipetting technologies for Freedom EVO workstations

To find out more on Tecan’s Air LiHa, visit www.tecan.com/airliha

5 ml disposable tips now available for LiHa Arm Tecan has further extended the flexibility of the Freedom EVO Liquid Handling (LiHa) Arm with the launch of a 5 ml disposable tip. Previously only available to Tecan customers working with fixed tips, the unrivalled pipetting volume range offered by the LiHa Arm can now be exploited by customers who require the highest levels of sample security.

The 5 ml LiHa disposable tips are available in either filtered or non-filtered options in Tecan Pure format, and are ideally suited to a wide range of applications in the pharmaceutical, academic and clinical sectors. The new tips can be used with

existing LiHa Arms, requiring only minor hardware modifications, and allow customers to take advantage of the complete range of LiHa disposable tips – from 10 μl to 5 ml sizes – using a single pipetting arm. This offers maximum throughput and flexibility for applications such as biobanking, large volume extractions and veterinary, food and clinical testing, allowing bulk liquid transfers to be rapidly performed without additional pipetting hardware.

To find out more on Tecan’s 5 ml LiHa disposable tips, visit www.tecan.com/consumables

Rack of 5 ml disposable tips for LiHa Arm

PRODUCT NEWS TECAN JOURNAL 1/2012

Tecan launches Infinite® M1000 PRO with AlphaScreen® and AlphaLISA® technology Uniformity

raw data

The Infinite M1000 PRO is Tecan’s latest generation high-end multimode microplate reader, packed with application-focused functions and technical innovations to offer exceptional speed and sensitivity for academic, biotechnology and pharmaceutical research.

Building on the success of the original premium Quad4 Monochromators™‑based Infinite M1000, the Infinite M1000 PRO provides greater flexibility than ever before, incorporating a range of user-oriented features to further enhance microplate-based assays.

The Infinite M1000 PRO is available with a module designed specifically for AlphaScreen and AlphaLISA technologies, taking advantage of the instrument’s high power laser light source to provide a comprehensive solution for this powerful bead-based assay system. The AlphaScreen module includes dedicated filters for both AlphaScreen and AlphaLISA assays, as well as an ingenious temperature correction function that can compensate for sample temperature variations across the microplate. This novel system helps to accelerate analysis of these

Infinite M1000 PRO

TC data

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Comparison of results with and without temperature correction in the Infinite M1000 PRO

highly temperature sensitive assays by eliminating the need for time-consuming equilibration of plates, increasing productivity without compromising on quality of results.

The Infinite M1000 PRO also offers excellent versatility for a wide range of other life science research applications, featuring patented technology for fine tuneable and precise fluorescence bandwidth adjustment and Tecan’s highly acclaimed absorbance, fluorescence and luminescence scanning capabilities. It incorporates a range of functions – such as Optimal Read (OR) and double orbital shaking – to accelerate and simplify experimental workflows and analysis, while maintaining the class-leading format flexibility and user-oriented features of the original Infinite M1000. The new system ensures

full compatibility with the latest laboratory techniques, offering absorbance, fluorescence, FRET, TRF, FP and scanning modes for a broad range of studies. It allows researchers to quickly switch between formats and assay types, offering pre- and user-defined microplate formats from 6- to 1,536-wells, as well as compatibility with Tecan’s patent pending NanoQuant Plate™ for low volume nucleic acid measurements, and can be fully integrated with the Freedom EVO liquid handling workstation for walkaway assay automation.

To learn more about the new Infinite M1000 PRO, visit www.tecan.com/infinitem1000pro AlphaScreen and AlphaLISA are registered trademarks of PerkinElmer, USA.

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PRODUCT NEWS TECAN JOURNAL 1/2012

Reinventing drug titration In the last issue of the Tecan Journal, we announced the availability of the revolutionary HP D300 Digital Dispenser exclusively through Tecan’s Life Science Business. As customers in Europe and North America begin to place their orders, Kevin Peters, HP Global Sales Development Manager, reports on early customer experiences with the new technology.

Titration is an everyday necessity for drug discovery biologists, and we have worked closely with customers to develop the product they need. For what might be considered a standard protocol, there is a surprising amount of diversity in how titrations are performed. This is because, until now, researchers have had to make a compromise, balancing quality of data with productivity, throughput and cost. Each laboratory makes a different trade-off, and so performs titration in surprisingly different ways. Early adopters of the HP D300 Digital Dispenser have been astounded at how digital titration technology has standardized this technique, radically improving quality, throughput and cost, as well as eliminating variability between operators. As a result, this technology has the potential to eliminate manual serial dilutions – and possibly even the serial dilution process – from drug discovery research.

As researchers become familiar with this technology, we are likely to see it being used for more and more applications, and there are already examples of how digital titration has enabled new science that was previously impractical; allowing study of drug-drug combinations as a potential avenue for new therapeutics, as well as providing access to some of the biological mechanisms involved in target binding for the first time. Our understanding of many of these interactions is being advanced simply by learning faster; you can now go from theory to experiment to results very quickly. Scientists are also able to perform more experiments with the same limited resources, allowing higher risk experiments to be performed earlier in the drug discovery process, potentially offering greater rewards.

The HP D300 allows direct dispensing to bioassay plates, for faster experimental set-ups and less waste

Overall, interest in the marketplace is very high, and the HP D300 has already won two awards. This early success is due to its combination of affordable technology – making it very attractive to low to medium throughput laboratories – and the system’s ability to quickly and conveniently standardize

processes for contract laboratories and large pharmaceutical customers. It really does offer something for everyone.

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

CORPORATE NEWS TECAN JOURNAL 1/2012

Customers benefit as Tecan Australia becomes a subsidiary Meet the team Customers in Australia, New Zealand and the Pacific Islands have welcomed the news that in early 2012 Tecan Australia became a fully owned Tecan subsidiary. A new facility in Melbourne and local presence in several major cities reflects Tecan’s uniquely strong commitment and continuing investment in the Australasian market. The new Melbourne base houses a comprehensive sales and support organization, complete with a workshop and additional facilities for customer training and demonstrations.

Especially important for customers spread over such a wide geographical territory, the new headquarters includes additional warehouse facilities and stock levels to quickly support customers in need.

Roland Durner, General Manager for Tecan Australia, said: “This is great news for our customers and employees alike.

Our new facility, backed by strong operating procedures and with Tecan’s renowned emphasis on regulatory and quality requirements, will help us to improve every element of our business. Our existing team of highly trained sales and support staff will now be even better placed to provide a dedicated service to our customers in research, clinical diagnostics, forensics, and food and agriculture in the region.”

To find contact information on Tecan Australia, visit www.tecan.com.au

Roland Durner

Luke Danielewski

Adam Gangur

James Hunter

Biju Kishor

John Ince

Clair Joyce

Josh Ardley

To hear more about Roland’s views on the challenges of the Australasian market, read this month’s Leading the debate on page 27.

Tecan Australia also provides a local presence at scientific meetings and exhibitions

Michael Zhen

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CORPORATE NEWS TECAN JOURNAL 1/2012

4th Tecan Symposium enjoys Chinese success Tecan recently hosted the fourth annual Tecan Symposium, welcoming leading genomics, proteomics and biobanking experts from around the globe to discuss this year’s topic; Translational Medicine: From bench to bedside and back.

Held outside of Europe for the first time, the 2011 Tecan Symposium was in Shanghai, China, reflecting Tecan’s strong commitment to the Asian market and the growing importance of China as a major center for genomics and clinical research. The two-day event was divided into four topics – current trends in translational research, clinical aspects of translational medicine, the growing importance of biobanking and the impact of technology on translational science – and explored a wide range of current hot topics in this fast-growing area of life science research.

Following a brief welcome from CEO Thomas Bachmann, the first day’s presentations started with a short retrospective on the field of genomics from Professor Huanming Yang, President of China’s very own

‘genome factory’, the Beijing Genomics Institute (BGI). Professor Yang highlighted how the publication of an organism’s genome leads to an increase in research on that organism, as well as the rapid development of genomics technologies since the beginning of the human genome project over 20 years ago. Continuing the technological theme, Dr Konstantinos Petritis from the Translational Genomics Research Institute (TGen) in Arizona, USA, discussed an innovative solution for protein biomarker verification and validation, stressing the importance of laboratory automation in achieving the required throughput and reproducibility. This was followed by Dr Mark Templin from the University of Tübingen, Germany, whose group is using protein microarray technologies to investigate potential disease and toxological markers,

Dr Michael Christman discussed the current and future roles of biobanks

CORPORATE NEWS TECAN JOURNAL 1/2012

Tecan CEO Thomas Bachmann welcomed delegates to the 4th Tecan Symposium

taking advantage of a wealth of new and archived histological specimens to accelerate biomarker discovery and validation. The session was concluded by Dr Kevin Rosenblatt from the University of Texas, USA, who presented data on a recent study into circulating biomarkers which could help to predict the risk of preterm birth, potentially helping to alleviate a major cause of infant morbidity around the world.

The second session kicked off with a live teleconference with Dr Paul Rhyne from Bristol-Myers Squibb, USA, who provided an insight into the practical considerations of using biomarkers in a clinical setting, and was followed by a presentation on the latest biomarker discovery applications of SUPRA and MALDI mass spectrometry techniques from Dr Patrick Ducoroy of the

Dr Kevin Rosenblatt talked about some promising biomarkers to predict the risk of preterm birth

CLinical Innovation Proteomic Platform (CLIPP), France. Mr Shailesh Maingi from the Clinical Trial Division of LabCorp then discussed the current challenges and possible future direction of personalized medicine, followed by some practical examples of how existing genomics knowledge is being put into practice for diagnostics, prognostics and epidemiology investigation from Dr Christopher Wong from the Genomics Institute of Singapore.

The second day of the Symposium concentrated on biobanking, and was begun

by Dr Michael Christman, President and CEO of the Coriell Institute for Medical Research, USA, one of the world’s largest biobanks.

Dr Christman looked at the current and likely future roles of biobanks in realizing the potential of genomics and proteomics

The Symposium also provided plenty of time for informal discussion of each day’s topics

discoveries, reminding delegates that, while the cost of sequencing a genome will soon be below $1000, the cost of interpreting that data is still millions of dollars. Dr Mark Cosentino from SAIC-Frederick, USA, then discussed the recent technological advances in biobanking itself, shattering the myth that a biobank is just a room full of freezers. Dr Mark Divers from the Karolinska Institute Biobank, Sweden, reinforced this message with a look at a number of collaborative research initiatives being undertaken by biobanks around the globe, highlighting the many and varied challenges that face these large multi-center collaborations. The session was concluded

by Dr Frank Rühli, who took a step back through time to show how ancient biobanks – mummified remains from thousands of years ago – still hold valuable information about the evolution of many diseases.

Many Tecan staff, including Dr Martin Brusdeilins (Tecan CSO, left) and Raymond Chan (Tecan Head of China and South East Asia, right), were on hand to talk to customers about the current needs in translational medicine

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CORPORATE NEWS TECAN JOURNAL 1/2012

The new HP D300 Digital Dispenser generated a lot of interest amongst delegates

The final session looked at some exciting recent technological advances, and was opened by Professor Yi Lu from the University of Illinois, USA, who showed how even well established technologies such as the near ubiquitous glucose meter can be used for novel diagnostic applications. This theme was carried on by Professor Changhuei Yang from the California Institute of Technology (Caltech), USA, who demonstrated how the power of another common technology – cell phone imaging sensor chips – could be used as an alternative to conventional microscopes for a range of cell culture studies. This was followed by a potential application of this novel imaging technology from Dr Tarik Issad, who described a BRET technique for the real‑time study of protein-protein interactions in living cells. The last speaker of the Symposium was Dr Colin Campbell from the University

Shanghai offered a unique cultural experience for delegates

of Edinburgh, who discussed an exciting joint project to identify the chickenpox- and shingles-causing Varicella‑zoster virus using protein microarrays. Rather fittingly, this collaboration was first conceived during a chance meeting at the very first Tecan Symposium in Zurich, highlighting the scientific value and excellent networking opportunities offered by this innovative event.

once again highlighting Tecan’s innovationdriven, customer‑focused approach.

To find out more about last year’s Tecan Symposium, visit www.tecan.com/symposium2011

Alongside the scientific content, Tecan’s local representatives – Tecan (Shanghai) Trading Company Ltd – arranged a number of social events for delegates, including a river cruise to provide a more relaxed environment to discuss the day’s topics, and a visit to a traditional Chinese tea house to celebrate the rich cultural heritage of Shanghai itself. Overall, the 4th Tecan Symposium was hailed a great success,

Dr Marina Lomtatidze (left) from Delrus Inc., Tecan’s distributor in Russia, and Professor Larisa Chernousova, Head of the Microbiology Department at the Central TB Research Institute of RAMS, enjoying the atmosphere at the tea house

A visit to a traditional Chinese tea house provided some local culture

QUALITY ASSURANCE TECAN JOURNAL 1/2012

China embraces a whole new dimension in blood safety Blood safety is a serious concern for countries worldwide and China is no exception to this.

For many years commercial blood donation has been prohibited in China, and the Government has adopted a systematic approach to continually improving blood quality, from the initial donation through to collection, transportation, screening and finally, transfusion in a hospital.

The Chinese Society of Blood Transfusion has a long association with Tecan, beginning in the early 1990s. Since the installation of China’s first modern liquid handling workstation at the Kunming Blood Center in Yunnan province, there has been close collaboration between the professional staff of the Society and Tecan, in a bid to steadily improve both the capabilities and the quality of blood screening services.

In addition to bringing the benefits of the latest instrumentation and automation to the Society, Tecan is keen to share its expertise and knowledge of quality control and quality assurance with the laboratories based at the Chinese blood centers and blood banks. In 2007, Günter Weisshaar, Senior Vice President of Tecan and Head of Quality Assurance and Regulatory Affairs, was invited to speak at the Annual Conference of the 16 blood centers and blood banks within Yunnan province, and gave a presentation on establishing a quality control system according to international quality standards. Günter has continued to address the conference every year since and, in 2011, was again invited to speak at this forum, raising the quality standard of the local blood screening facilities to new heights when he introduced the concept of enterprise risk management and its impact on blood safety assurance. Enterprise risk management systems ensure that blood centers and blood banks are prepared for any unexpected disaster that could lead to a breakdown in their ability to process blood. Using all the knowledge and experience gained by Tecan during the implementation of its own enterprise risk management system, Günter systematically described the framework, methodology and documentation required.

The clear goal was to advise delegates on

Günter Weisshaar presenting on enterprise risk management

the possible risks and how they could be identified, evaluated and mitigated.

The presentation triggered a tremendous discussion among the delegates at the conference. In many cases it was not the first time they had heard about risk management, but being shown a systematic and comprehensive approach was clearly quite enlightening and some delegates were motivated to express an interest in implementing their own scheme, requesting Tecan’s advice and assistance. Tonghan Yang, Director of the Kunming Blood Center, commented: “As our standard operating procedures and quality control systems mature, the focus of our attention to further improve blood safety is now shifting to risk management, and Tecan’s presentation was both timely and very helpful. We are determined to continuously build on our

capabilities and are planning to initiate a risk management scheme at the Kunming Blood Center early in 2012. Tecan’s helping hand to guide us through this journey will be greatly appreciated.”

Mrs Pu Yang, Director of Hong He Gu blood bank, expresses an interest in implementing an enterprise risk management scheme in her organization

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CELL BIOLOGY TECAN JOURNAL 1/2012

Successful isolation and expansion of human primary cells A network project jointly funded by the Swiss Confederation CTI (Commission for Technology and Innovation) and Tecan, bringing together industrial, academic and clinical partners such as the Zurich University of Applied Sciences (ZHAW) and the University Hospital Balgrist in Zurich, has developed a successful automated system based on the Freedom EVO® platform. The system isolates and cultivates cells from human intervertebral disks, improving reproducibility and encompassing all-­important integral quality control.

(Medic Tool, Switzerland). Incorporating this early stage of sample preparation into the automated workflow was a main goal and aimed to avoid inconsistencies in tissue handling by different laboratory personnel.

The homogenized tissue was then further digested and washed to obtain purified single disk cells. Finally, cells were seeded into Corning RoboFlasks® using proliferation medium. To ensure the optimal conditions for cell proliferation, the medium was changed twice a week and, when about 70 % confluence was reached (typically after 5-6 days), cells were harvested and re-seeded.

The Cell Biology and Tissue Engineering Division at ZHAW. Front row left to right: Ursula Graf­-Hausner, Epifania Bono, Stephanie Mathes, Marta Kley; back row left to right: Nicola Franscini, Diego Santini

As more laboratories around the world target the growth of human cells for applications in tissue engineering, there is an urgent need for improved consistency in cell culture processes and automation is playing an increasingly important role. Professor Ursula Graf-Hausner, Group Leader of the Cell Biology and Tissue Engineering Division at ZHAW, explained: “Tissue engineering for regenerative medicine is a challenging and promising technology, but it is expensive and many hurdles must be overcome before the technology reaches the clinical arena. In 2007, my clinical colleague Professor Norbert Boos and I first approached Tecan as an industrial partner to look at the advantages automation might bring to cell-based technologies, with the long term aim of making them more standardized and, effectively, more suited to clinical applications.”

A fully automated procedure was established on a Freedom EVO 150 workstation, configured using typical modules for automated cell culture. Freedom EVOware® was used for script generation and device control. All steps from enzymatic isolation of cells to proliferation of the isolated cells – including harvesting and seeding – and on-system quality control were automated.

Parameters such as yield of isolated cells, viability, aggregation and growth rates, as well as phenotype, were analyzed to compare the automated and manual procedures.

The procedure used biopsy samples taken from patients undergoing spinal surgery (disk herniation). In the manual procedure, tissue was cut into small fragments whereas in the automated procedure, the tissue was homogenized using the Dispomix® system

In the manual procedure, regular determination of cell number was carried out after trypsination using an image-based Cedex™ Analyzer (Roche) for cell counting.

Confluency measurement was implemented in the automated method using an integrated Roche Cellavista™ System. The Cellavista System is non-­invasive and can be used to accommodate different growth kinetics for different donors. This is a huge advantage, since cells from one patient may take longer to grow than those from another patient. Another real benefit of the system was the ability to predict the number of cells expected at a certain time; by calibrating the system with specific cell growth parameters, the team was able to non-­invasively correlate confluence with an expected cell number after harvest. This online monitoring also served as a warning option if predicted growth curves did not correspond to the measured values.

Establishing intensive quality control at each step and comparing the manual and

CELL BIOLOGY TECAN JOURNAL 1/2012

automated processes were absolutely essential for the project to succeed.

Phenotype comparison was achieved by immunostaining (collagen I, collagen II, versican and total proteoglycan) of cells grown on glass slides for two days, either manually or on the Freedom EVO using the same staining protocol. Immunofluorescent analysis was performed using the Cellavista System (automated procedure) or an Axioskop® 2 Plus microscope (Zeiss) manually.

The analysis of isolated primary cells and of cellular characteristics demonstrated that the automated process resulted in the same number, viability, phenotype and even growth rate of cells as the manual method.

Overall, the procedure allowed successful isolation of human intervertebral disk cells from biopsy and their expansion over several passages. Professor Graf-­Hausner concluded: “The automated system achieved all our aims; we achieved excellent levels of reproducibility and standardization, which is essential when you’re working with human material that could potentially be used therapeutically. From the technical and scientific point of view, the challenges of cell-based therapy have effectively been solved; the next step will be to look at these techniques in animals and then pre-clinical studies. We continue to have an excellent collaborative working relationship with Tecan, which is essential for success because we are biologists and not equipment experts.

We have several small projects underway and the Freedom EVO is so flexible that we can adapt it depending on the questions we want to answer.”

Full details of this study can be found in: Franscini, N. et al. J Lab Autom, 2011, 16(3), 204-13.

For more information on the Zurich University of Applied Sciences (ZHAW), visit www.zhaw.ch/en.html

To find out more on Tecan’s Freedom EVO liquid handling workstations, visit www.tecan.com/freedomevo

Dispomix is a registered trademark of Miltenyi Biotec.

RoboFlask is a registered trademark of Corning.

Cellavista and Cedex are trademarks of Roche.

Axioskop is a registered trademark of Carl Zeiss AG.

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4 1. LiHa Arm with four standard tips and four wide bore tips for cell pipetting 2. Robotic Manipulator (RoMa) Arm equipped with eccentric gripper fingers 3. Flipper module with four positions for RoboFlasks 4. Centrifuge Rotanta 46 RSC Robotic (Hettich, Germany), mounted under the worktable 5. Clean air hood 6. Cellavista System 7. Orbital shakers Not shown: Optional incubator for fully automated system (37 ˚C, 5 % CO2, humidified atmosphere)

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GENOMICS TECAN JOURNAL 1/2012

Biobanking clinical cytology samples in Sweden The Swedish national biobanking infrastructure, BBMRI.se (BioBanking and Molecular Resource Infrastructure of Sweden), has initiated a pilot study for the improvement and national harmonization of biobanking procedures in clinical cytology.

Following a national procurement process, the first of 10 Freedom EVO® liquid handling platforms has been installed in the Clinical Cytology Biobank at the Karolinska University Hospital.

Nasrin Perskvist loading samples onto the Freedom EVO workstation

ThinPrep® Pap Test containers can be loaded straight onto the Freedom EVO

The Karolinska University Hospital at Huddinge, Sweden, is launching a new cervical screening algorithm that includes testing for human papillomavirus (HPV) persistence by testing for the virus in biobanked samples from previous screening rounds. HPV is a known cause of cervical cancer, and the use of HPV testing in cervical screening has long been under consideration.

However, as many HPV infections clear spontaneously, it is necessary to know whether an infection is new or long-standing.

Systematic biobanking will allow this new screening initiative to be introduced, and will also allow thorough comparative evaluation

GENOMICS TECAN JOURNAL 1/2012

Processing of 96 samples on the Freedom EVO workstation

Sample tubes automatically decapped and 4 ml aliquots transferred to intermediate tubes

Storage plate transferred to freezer

Tube recapped

Offline sealing of storage vials

Patient samples arrive in laboratory in ThinPrep containers

ThinPrep containers loaded onto Freedom EVO and identified by barcode scanner

240 μl aliquots aspirated from bottom of tube (layer containing most cells) and dispensed into individually barcoded 300 μl vials in 96-tube storage plate

Sample allowed to sediment for 30 minutes

Summary of the Karolinska’s current workflow

of the many new cervical screening tests that have recently been marketed.

In Sweden, cervical screening takes place every three to five years between the ages of 23-60, primarily using cytological procedures followed by HPV testing. Karolinska is now introducing an algorithm that instead initially performs HPV analysis, as has also been introduced in the Netherlands and Mexico.

Nasrin Perskvist, Project Coordinator, explained: “Because of the importance of cervical cancer screening, the biobanking initiative will provide a vital resource for improvement and quality assurance of

screening protocols and tests. These cells may also be useful for basic research on cervical cancer, as well as on other diseases that affect women. By launching a national cytology biobanking initiative we aim to provide a resource that improves both research and patient healthcare.”

“We wanted to build a biobank using the cell samples from Sweden’s national cervical cancer screening program. Liquid-based cytology is generally useful for both diagnostics and subsequent research, and storing these samples in a quality assured system offers many benefits for both

researchers and patients.” The project began in 2010, with Nasrin investigating temperature, cell concentration and other factors important for sample storage. Cervical cells are collected into PreservCyt®, a methanol-based medium, and can be stored at -20 to -25 °C without freezing. This is a significant advantage in cytodiagnostics, as slides can be made repeatedly – using different staining methods if required – without freeze-thaw cycling of the sample, maintaining good cell morphology. These cells are also a very good source of DNA and RNA for HPV testing or other biomarker analysis.

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GENOMICS TECAN JOURNAL 1/2012

With support from the BBMRI.se, the Karolinska Hospital now has an automated method to provide good quality assurance and enable long-term storage of these samples. Nasrin continued: “We defined our specifications and then put out an invitation to tender. Several companies showed interest and, after careful consideration, we chose Tecan. We visited Tecan on several occasions to discuss and finalize the development and validation of the system in collaboration. We optimized the process manually, and within three months Tecan had adapted it to an automated system tailored to our needs. The system is based on a Freedom EVO® 150 liquid handling platform, equipped with 2­-channel Liquid Handling (LiHa) and Pick and Place (PnP) Arms, a customized decapper and a barcode reader to provide full traceability of all samples. Final modifications – to accommodate real patient samples in ThinPrep® containers – and system optimization were performed on site.”

Barcoded sample jars are moved to the decapper by the PnP Arm, and 4 ml of each sample is transferred to its corresponding intermediate tube by the LiHa. After a 30 minute sedimentation period – which is just as effective as centrifugation and fits nicely into the workflow with no periods of inactivity – a 240 µl aliquot of cells is transferred to the final storage plate, which contains 96 individually barcoded 300 µl vials. The barcodes are compatible with both the automation software and the LIMS, providing full sample tracking throughout the entire workflow. The final storage plate location is identified by the LIMS, which controls a spacious, electronic freezer and will ultimately

link all the collaborating cytology laboratories that will eventually use the BBMRI.se system.

Nasrin added: “Over the past few weeks, we have been able to run between 350 and 400 samples a day without any difficulty; a run of 96 samples takes 2 hours 40 minutes, which we are very happy with.”

Karolinska University Hospital is the first site to trial the system which, once established, will be installed at other sites throughout Sweden. Almost 700,000 samples are collected annually across the whole of the country and, at present, 80 % of Sweden’s national cervical screening program uses liquid-based sampling that is suitable for biobanking. In Stockholm – where some 100,000 samples are taken each year – there are three different cytology laboratories which all supply the biobank, and about 10,000 samples are already in storage, although it is too soon for any of them to have been used. “We need to announce the availability of this resource to all the relevant healthcare professionals and researchers, but by late summer/autumn 2012 the connections will be established. Everything has been developed in accordance with national regulations and ethical standards, and the beauty of the biobank is that it will benefit not only the cytology laboratories and the researchers, but also the patients themselves,” said Nasrin. “Working with Tecan to develop this custom solution was very exciting. The joint expertise of BBMRI.se and Tecan has enabled us to achieve our overall aim of cost-efficient and high quality biobanking of these very precious samples.”

To find out more on Tecan’s Freedom EVO workstation, visit www.tecan.com/freedomevo To find out more about the Karolinska Institute, visit www.ki.se To find out more about Karolinska University Hospital, visit www.karolinska.se/en To find out more about BBMRI Sweden, visit www.bbmri.se ThinPrep and PreservCyt are registered trademarks of Hologic Inc. and/or its subsidiaries in the United States and/or other countries.

services TECAN JOURNAL 1/2012

Braving the elements in Norway with Te-Care™: a complete service package for better patient care CO

Medical Technologist Mette Bogen using the FE500pro

“Instrument reliability and service support were key considerations in this decision, as many companies are not able to provide adequate technical assistance to meet the needs of clinical laboratories in Scandinavia, and we knew that Tecan would be able to provide the level of support we required. Our system is covered by a Te-Care Complete service contract, which includes both routine maintenance and technical assistance in the event of a problem, giving us complete peace of mind.”

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Vestre Viken Hospital Trust serves a population of over 450,000 people in and around Drammen. The Trust’s main laboratory, situated at Drammen Hospital, performs over three million analytical tests a year, and has fully automated its front-end sample processing using an FE500pro pre-analytical sample handling platform. Ragnhild Grondahl, Medical Technologist, explained: “Our laboratory performs a wide range of clinical tests, serving both the hospital and a large number of primary care centers in the area.

In 2005, to help us deal with an ever increasing workload, we decided to automate our pre-analytical sample handling, and looked at a number of instruments before choosing an FE500pro system from Tecan.”

MPL E TE

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Drammen Hospital, Norway, relies on a Te-Care Complete service contract to ensure reliable operation of its FE500pro™ pre-analytical sample handling platform, helping the laboratory to provide clinical analysis services for both the hospital and local primary care centers.

“The Te-Care contract includes regularly scheduled maintenance, and this is essential in preventing problems from arising. This is particularly important in more remote markets, such as Norway, as we cannot afford lengthy periods of instrument downtime without impacting on patient care. This approach means that we have had very few issues with the platform, but we have always had a good relationship with Tecan service personnel – first in Switzerland and more recently here in Norway – if we do need help. Many problems can be quickly fixed over the phone, and Tecan’s engineers are good at explaining technical procedures in terms that are easy for medical technicians to understand. This not only minimizes downtime, it also means that our staff become more knowledgeable and competent with the instrument, making them feel more confident and empowered to use it effectively. Overall, we are very happy with the level of service we receive, safe in the knowledge that support is always available if we need it.”

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

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GENOMICS TECAN JOURNAL 1/2012

Making sweet music: art and science join forces for the LabNotes project

LabNotes, a unique crossdisciplinary project that emerged from the Scottish Crucible program, will see academics from the diverse fields of systems biology, science communication and music working together to portray science in a radically different way by producing a musical film of a Freedom EVO® workstation in action.

From left to right: Martin Clark, Jane Stanley, Eliane Salvo-Chirnside and Lorraine Kerr on the first filming day of the LabNotes project

The Scottish Crucible is a highly successful professional and personal leadership and development program based on a model set up by NESTA, the UK’s National Endowment for Science, Technology and the Arts. The program aims to train potential research leaders of the future and promote interdisciplinary collaboration across the Scottish research community. During the course of the 2011 program, scientist Dr Lorraine Kerr1 and composer Dr Jane Stanley2 began to formulate a collaborative project to create a novel musical composition, with an accompanying short film, focusing on the Freedom EVO liquid handling workstation. Joined by Dr Christine Knight3, a specialist in science communication, and freelance film maker Martin Clark 4, the team applied for and were awarded a grant to put their ideas into action.

The Freedom EVO that is the focus of this extraordinary project normally serves as a high throughput systems biology instrument at the Kinetic Parameter Facility (KPF) in the Centre for Systems Biology, the School of Biological Sciences, at the University of Edinburgh.

Dr Kerr manages access, throughput and protocols for the Freedom EVO on a daily basis, and explained what the LabNotes project hopes to achieve: “Currently in the science community there is a real push for public engagement of your science and this seems like an interesting way to look at what we do with a completely novel approach.”

Filming for LabNotes has begun, recording the robot’s movements in the course of a

normal everyday script with changing angles and lighting, and musical composition will begin once this is completed. A launch event for the final piece is planned for 2012, after which it will be published on several online sites and submitted to science and film festivals. This project will undoubtedly provide a completely different viewpoint on how to engage with the public and explain science in a captivating way – Tecan looks forward to seeing the results!

Don’t give up the day job!

The Freedom EVO is used virtually all day every day by a number of different groups; the KPF is a hub of substantial life science equipment and expertise available to researchers aiming to establish mathematical models of biological systems in a range of organisms. Dr Kerr added: “We continually push technologies to their limits, simply because that’s what academia is all about, and this was very much a consideration when we first chose the Freedom EVO workstation as our high throughput automation platform.

We knew our workload would comprise a significant number of quantitative PCR assays but we also needed the flexibility to change, for example, to automating RNA extraction, if required. As academics, we wanted the freedom to modify configurations of the workstation occasionally, largely because our center works on many different organisms, from plants and yeast, to E. coli

GENOMICS TECAN JOURNAL 1/2012

and mammalian cells; we wanted to be able to do this without being tied to specific chemistries. We presented very specific requirements in the form of a European tender and Tecan had the edge over its competitors on the versatility and reliability of the system, plus the ease with which we could develop our own scripts once we were familiar with the software.”

The wide range of pipetting volumes possible on the Freedom EVO was also an important advantage; it is capable of pipetting 1 μl very accurately, with minimal risk of carryover, but still has the flexibility to pipette milliliters of liquid if necessary. “We have calibrated our system to accurately aspirate and dispense 1 μl and test it every six months using gravimetric testing. For most experiments we use fixed tips and a TipCharger™ (IonField Systems™) which, in my experience, is quite unusual for qPCR. We initially tested this set-up by pipetting a million copies of a gene and were impressed to see absolutely no carryover. Similarly, we have thoroughly tested how the system handles RNA for one-step real-time PCR because RNA is very unstable, and found that there is absolutely no problem; the RNA does not degrade at all and it is very clean.”

Dr Kerr and her team recently automated RNA extraction from plants, which is renowned to be very difficult, and a paper has just been published in Plant Methods (Salvo‑Chirnside et al, Plant Methods 2011, 7:40).

Martin Clark takes some close-up shots of the Freedom EVO in action Please note, the top window panel of the Freedom EVO has been removed for filming and the instrument is switched off

To find out more about Tecan’s genomics applications on the Freedom EVO, visit www.tecan.com/genomics To find out more about the Scottish Crucible, visit www.hw.ac.uk/scottishcrucible 1 Dr Lorraine Kerr, Project Manager for Experiments, Centre for Systems Biology at Edinburgh (CSBE), School of Biological Sciences, the University of Edinburgh. www.csbe.ed.ac.uk/people/profile/lorraine-kerr

Dr Jane Stanley, Lecturer in Music, School of Culture and Creative Arts, the University of Glasgow. www.janestanley.com/About.html 2

Dr Christine Knight, Genomics Forum Policy Research Fellow, ESRC Genomics Policy and Research Forum, the University of Edinburgh. www.genomicsnetwork.ac.uk/forum/people/ academicstaff/forename,3660,en.html

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4 Martin Clark, freelance film maker. www.martinrmclark.com/statements_amp_cv

TipCharger and IonField Systems are trademarks of IonField Systems, Delaware, USA.

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readers TECAN JOURNAL 1/2012

Unwinding the genetic clock Researchers at the Institute of Entomology (ENTU) in the Czech Republic are studying insect circadian clocks and have used Tecan’s Infinite® M1000 plate reader to automate hourly luminescence measurement of transgenic Drosophila. Automation enables large-scale screens of mutant Drosophila strains to be performed, involving hundreds of measurements over a period of up to two weeks.

Scientists at the Laboratory of Molecular Chronobiology in the Department of Molecular Biology and Genetics, Institute of Entomology, České Budějovice, are studying the circadian clock in Drosophila melanogaster, the fruit fly, to better understand the cellular and molecular mechanisms underlying internal rhythms.

Dr David Doležel, a researcher in the Laboratory of Molecular Chronobiology, explained: “Insects are very convenient for genetic research, and studying Drosophila is valuable for mammalian research. Like humans, insects have an internal clock so that they know whether it is morning or evening, and the circadian genes – which are essential for this clock – are quite conserved between insects and humans. To study the expression of these genes, which are cyclically expressed over a 24-hour period, we have created transgenic Drosophila containing luciferase reporter genes controlled by the promoter of the circadian gene under study.

Individual flies are placed into 96-well microplates, one fly per well, and surrounded by agar containing food, water and luciferin.

We then measure the luminescence caused by activation of their reporter genes at hourly intervals for up to ten days.”

“To reliably perform this assay on a large scale, we needed a microplate reader that would enable us to automate plate transfer; this is essential for our research as measurements are performed at regular intervals throughout the day and night. We also wanted an instrument with a luminescence module that was equipped with filters, as this would allow us to discriminate between red and green luciferases, enabling us to follow multiple genes per well. Tecan’s Infinite M1000 plate

reader satisfied all our requirements; it can be used with a variety of microplate formats, it is available with luminescence color filters and it is equipped with a reliable stacker system which automatically transports the microplates to and from the reader. Its compact size also means that it can be fitted into our laboratory incubator, allowing us to precisely control light and temperature and investigate the effect of environmental variables on the regulation of the circadian gene being studied.”

David continued: “The Infinite M1000’s microplate format enables us to load hundreds of flies into the plate reader at the same time. The bioluminescence emitted by the living flies is then measured at hourly intervals for up to two weeks, providing us with a fully automated system to screen for mutations that affect circadian gene activity and to test new variations that we have created in house. In a typical experiment,

Bioluminescence (CPS)

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we measure bioluminescence from around 500-600 flies in eight plates, usually four to six individuals per mutant strain. To ensure light can access the flies, we alternate empty transparent plates with those containing flies, and in one month we can screen for between 200 and 400 different mutant strains. Of those strains, 99 % will be discarded after the two week screen, only retaining the mutants of interest to create new mutations for further study.”

“We also use Drosophila tissue cultures or isolated organs to study circadian gene expression through luminescence, and these too are in 96-well format for the luciferase assay. This format is useful for developing new assays, and we have already successfully been able to independently measure the luminescence of red and green luciferases in Drosophila Schneider 2 cells, thanks to the Infinite M1000’s luminescence filter system.

This strategy allows us to follow multiple

Bioluminescence: example of bioluminescence recorded from individual transgenic Drosophila melanogaster

Time (days) In this experiment, a promoter of the circadian gene timeless drives luciferase expression in a temporal pattern identical to timeless mRNA, enabling scientists to trace periodic expression in individual flies for two weeks. The black and white bars under the panel indicate when lights were off (12 hr) and on (12 hr), respectively. Gray bars indicate subjective daytime in darkness (12 hr). (Dolezelova and DoleŽel, unpublished)

readers TECAN JOURNAL 1/2012

D. melanogaster (left), classical model of genetics compared to the housefly, Musca domestica

target genes in a single assay, which we can hopefully extend to living flies in the future.

The Infinite M1000 can also be used with other formats – such as 24-well microplates – which, while not used in our current research, may be useful for other assays in the future; it is good to know that this option is available to us.”

“We have been very impressed by both the performance and the flexibility of the Infinite M1000, and Tecan was very happy to allow us to visit its premises in Austria

to test the system with our own samples before we committed ourselves to purchasing the instrument. We spent an entire day testing the reader for our assay, and then Tecan’s own engineers further developed the instrument’s stacker system to meet our requirements. We were delighted to have this opportunity to trial the plate reader to see how it would handle our Drosophila cultures.

Genetic research on insects requires the use of huge numbers of these organisms, and the Infinite M1000 reader makes this large-scale screening possible,” David concluded.

To find out more about Tecan’s Infinite M1000 plate reader, visit www.tecan.com/infinitem1000 For more information on the Institute of Entomology, visit www.entu.cas.cz/en/

Recent technologies allow genetic modification of new model organisms, in this example the housefly, Musca domestica. The Green Fluorescent Protein (GFP) expressed in the eyes was used as a marker to trace the presence of a transgene delivered by piggyBac transposon. Three transgenic and one control fly were photographed under white light (left) and GFP fluorescence after blue light excitation (right). (DoleŽel, Hediger & Bopp, unpublished)

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CELL BIOLOGY TECAN JOURNAL 1/2012

A new dimension in plate preparation for 3D cell assays

Scientists at the AstraZeneca Innovation Center China have developed a novel technique using a Freedom EVO® system for temperature-dependent automated preparation of cell culture plates for candidate compound screening. By pre­ coating microplate wells with agar prior to pipetting the BD Matrigel™, the team is able to perform 3D cell-based assays in a 96-well microplate format.

AstraZeneca Innovation Center China

The Innovation Center China (ICC) is an AstraZeneca research and development facility in the Zhangjiang Hi-tech Park, Shanghai.

Opened in 2007, the Center specializes in the investigation of diseases affecting the Asian population, including gastric, liver and lung cancers. State­-of-the-art technology is used to elucidate the genetic mechanisms of these cancers and identify clinically relevant biomarkers and drug targets.

The Center’s Cell Science Discipline, led

by Dr Yi Gu, runs an active drug discovery program targeted towards these diseases, using a range of compound screening techniques to identify novel drug candidates.

One of the secondary screening techniques used by the group is a 3D cell-based assay where cells are grown on a BD Matrigel substrate (BD Biosciences). Matrigel is a gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. Matrigel induces the growth of colonies of cells in a different manner to the standard cell culture surface (Figure 1). Its heterogeneous composition stimulates the formation of colonies, making it ideally suited to the study of tumor cell-cell contact, growth, migration and metastasis. Since the colonies mimic the growth and formation of tissues in the body, they give the researchers the potential to identify the right compound in the very early phase of the drug discovery process.

Manual plate generation for the Matrigel assay is both problematic and time consuming, and so automation of the process with the Freedom EVO workstation was an obvious solution in order to increase throughput and improve consistency.

The platform was already being used for

compound serial dilutions and a number of other cell-based assays. It could quickly and economically be modified for this protocol without interfering with other applications.

William Shi, Senior Associate Automation Scientist at the ICC, explained: “Historically, these assays have been performed in 6-well plates. Because of the large volumes needed and the complex handling requirements of the Matrigel, the manual production of the plates was rather difficult. Additionally, this format is poorly suited to drug screening due to the large volumes of reagents and candidate compounds required, and so we looked at ways to optimize this assay for cost effectiveness. More importantly, Matrigel is extremely difficult to pipette in low volumes, as its heterogeneous composition leads to high surface tensions and uneven distribution across a well, making analysis of results extremely difficult, and impossible for 96-well plate formats. To overcome this, we developed a protocol to pre­coat the microplate wells with a thin layer of soft agar prior to adding the Matrigel. This solved the surface tension issues associated with the Matrigel, but further complicated pipetting logistics as agar solidifies below 50 °C and Matrigel becomes too viscous to pipette above 4 °C.” The Freedom EVO platform was then adapted for the assay plate preparation by addition of several temperature-controlled carriers, allowing both the agar and Matrigel reservoirs to be kept at their appropriate temperatures to allow pipetting. All pipetting is carried out using a single channel on the system’s Liquid Handling (LiHa) Arm, which is pre-heated or pre-cooled by multiple

CELL BIOLOGY TECAN JOURNAL 1/2012

aspiration and dispense cycles to avoid the risk of tip blockages. Initially, hot agar (at around 55 °C) is pipetted into pre-warmed 96-well plates using the LiHa Arm’s multi‑dispense function. This minimizes the risk of premature cooling of the media and allows small media reservoirs to be used, helping to further reduce costs. The plate then cools at room temperature on the deck of the instrument for 15 to 20 minutes, allowing the agar to solidify prior to addition of the Matrigel.

Using this new protocol, six complete plates could be prepared in one and a half hours, compared with only three or four plates in an entire working day when performed manually.

The quality of the plates has also improved tremendously using the automated technique, avoiding the batch­-to­-batch, plate-to-plate and even well­-to­-well variation that often occurs with manual plate preparation. William concluded: “This consistency is vital for drug screening applications, providing high quality results to help us accelerate the drug discovery process, and the flexibility of the Freedom EVO platform made this easy to achieve.”

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The Cell Science Discipline team. From left to right: Jenny Xia, William Shi, Bin Xiang, Yi Gu

To find out more on Tecan’s Freedom EVO liquid handling workstations, visit www.tecan.com/freedomevo

BD and Matrigel are trademarks of Becton, Dickinson and Company.

Corning is a registered trademark of Corning Incorporated.

To find out more about the AstraZeneca Innovation Center China, visit en.astrazeneca.com.cn

B

Figure 1: Gastric cancer cell line SNU-668 colonies growing in Corning® 96-well clear flat bottom TC-treated microplate, A) without Matrigel B) on Matrigel

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CELL BIOLOGY TECAN JOURNAL 1/2012

High throughput hybridoma generation Tecan’s Freedom EVO® liquid handling platform has enabled Nurex to enhance its production of monoclonal antibodies, improving efficiency and increasing the number of clones obtained from each fusion.

From left to right: Dr Franca Mannu, Dr Marcella Orrù and Dr Franco Carta

Nurex is a biotechnology company based in Sassari and Turin, Italy, that specializes in the production of advanced diagnostic tools for biomedical applications in genomics, proteomics and clinical diagnostics. Dr Franco Carta, a researcher at Nurex, explained: “Nurex was established in 1995, developing and producing diagnostic kits for molecular biology applications such as the diagnosis of viral infections and some genetic mutations.

Ten years on, we expanded the range to include products and services for proteomics, before introducing the antibody production service in 2007. We mainly supply these antibodies to private research groups or university researchers, and initially used classical manual techniques to produce monoclonal antibodies. As the demand for antibodies grew, we needed to increase production and turned to automation to achieve this.”

During the evaluation phase, Nurex visited several other laboratories that use liquid handling workstations from various vendors to manage cell culturing and hybridoma generation. “We were impressed by the high flexibility of the Tecan platforms that would allow us to implement a wide range of protocols, and the high throughput they achieve in order to reduce manual operation.

We now have a Freedom EVO 200 platform equipped with a four-channel Liquid Handling (LiHa) Arm, a Robotic Manipulator (RoMa) Arm

and a MultiChannel Arm™ (MCA 96).”

The configuration also includes a microplate hotel, an incubator and two heated Te‑Shake™ modules for 50 ml tubes, to allow Nurex to meet the demand for increasing numbers of monoclonal antibodies. “The similarity of the language we already use for basic programming to that in Tecan’s Freedom EVOware® software made the transfer from manual processing to automation much easier,” Franco added.

Using the Freedom EVO liquid handling platform, Nurex has developed an innovative, high throughput methodology for monoclonal antibody production that allows hybridoma generation and cell culture maintenance to be performed in a sterile environment, with minimal user intervention to reduce the risk of errors and contamination. Automation ensures that a constant temperature is maintained throughout processing, which improves the quality of the results, and the LiHa guarantees rapid dispensing of consistent reagent volumes of media and buffers. The cellular fusion between a myeloma cell line and spleen cells from an immunized mouse is initiated by adding the required reagents while shaking and heating. The cells are then washed manually and the MCA 96 is used to transfer them to multi-well plates for incubation at 37 °C.

Monday

Tuesday

Wednesday

Thursday

Friday

Medium change

Fusion

Dot Blot

Positive hybridoma transfer

Positive hybridoma transfer

8 x 16 plates 13 day old fusions

8 fusions

8 x 16 plates 29 day old fusions

30 day old fusions

37, 54, 61, 68 day old fusions

Dispensing

Dot Blot

8 x 16 plates

36, 53, 60, 67 day old fusions

Schedule for high throughput monoclonal antibody production

CELL BIOLOGY TECAN JOURNAL 1/2012

Lymphocytes from immunized mice

Myeloma cell culture

Cell fusion and dispensing (16 x 96-well plates) Day 0 Medium change Day 13

Freedom EVO Dot Blot assay Days 29, 36, 53, 60, 67

Hybridoma transfer Days 30, 37, 54, 61, 68 Flask culture

Flow diagram of methodology for high throughput monoclonal antibody production

The culture medium is completely replaced after 13 days – the MCA 96 plays a key role in this – and a semi-automated Dot Blot assay is used to screen positive clones after 28 days.

For this assay, Nurex uses the MCA 96 and a third party pin tool (V&P Scientific) to transfer 100 nl of the culture supernatant from up to 16 fusion plates onto a Hybond™ ECL™ nitrocellulose membrane saturated with antigen, generating a total of 1,536 dots for analysis. The membrane is then incubated with a dye-conjugated secondary antibody, and washed before identification of positive colonies using an Odyssey® infrared scanner (LI-COR Biosciences).

Based on the results from this assay, the Freedom EVO workstation is programmed to transfer the positive cultures from 96- to 48-well plates. The assay and the analysis is repeated on a weekly basis for four weeks and, on each occasion, the positive hybridomas are transferred to another plate format, from 48- to 24-­well, 24- to 12-well, and so on. This methodology is at least 100 times more sensitive than ELISA, offering higher accuracy and selectivity, enabling generation of four or five monoclonal antibodies per fusion. In 2010, for example, Nurex performed over 300 fusions. This would be an impossible task if working manually, as only two fusions per day could be handled by manual processes.

Automation has enabled Nurex to increase the throughput four-fold, and has eliminated the variable fusion times related to the poor temperature control associated with manual techniques.

“The Freedom EVO workstation has proved robust and easy to use for cell culture, allowing us to automate all our protocols, defining and standardizing all the parameters and generating more reproducible results. All

fusions now result in hybridoma production, whereas with manual techniques this was not always the case, and our quality control procedures – the routine assessment of affinity and specificity – are more accurate,” concluded Franco.

To find out more on Tecan’s Freedom EVO, visit www.tecan.com/freedomevo To learn more about Nurex, visit www.nurexbiotech.com Hybond and ECL are trademarks of GE Healthcare.

Odyssey is a registered trademark of LI-COR Biosciences.

Antibody purification

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Taking special care of dangerous pathogens

The P4 Inserm-Jean Mérieux laboratory in Lyon relies on the flexibility, robustness and reliability of Tecan’s Infinite® 200 PRO monochromator-based microplate reader for absorbance-, luminescenceand fluorescence-based assays on Risk Group 4 pathogens.

The P4 Inserm-Jean Mérieux laboratory in Lyon is a highly secure laboratory, the largest of its category in Europe, affiliated to the French National Institute of Health and Medical Research (INSERM), and open to national and international scientific teams that need to handle Risk Group 4 pathogens. P4’s main activities include diagnosis, research, biosafety and training on agents responsible for viral hemorrhagic fevers (Ebola, Marburg, Crimean-Congo hemorrhagic fever and Lassa) and fatal encephalitis (Nipah and Hendra).

In his role as Biological Technics Engineer in the in vitro experimentation and diagnostic team at P4, Stephane Mely chose an

Researcher at the P4 laboratory

Infinite 200 PRO microplate reader for the many assays performed under Biosafety Level 4 (BSL4) conditions. He explained: “I am responsible for all scientific equipment, from purchasing to its final implementation in scientific programs and technical training of my fellow P4 scientists. We chose the Infinite 200 PRO specifically for absorbance-, luminescence- and fluorescence-based assays performed in accordance with very special BSL4 conditions. These include ELISAs, bioluminescence detection using luciferase activity, and pathogen detection with molecules coupled with fluorophores. As an example, we recently ran a fluorimetric assay looking at the neuraminidase activity of the influenza virus

TALK TO TECAN TECAN JOURNAL 1/2012

after reaction with new specific molecular inhibitors. Neuraminidase (sialidase) activity was detected by hydrolysis of the 4-MUNANA substrate (4-methylumbelliferyα-D-N-acetylneuraminic acid) into 4-methylumbelliferone. This hydrolysis ensures a shift of fluorescence spectra from substrate to product, and can be read at excitation 365 nm/emission 450 nm.”

One of the most important aspects of equipment in a Category 4 laboratory is robustness, not least because of limited access to the laboratory for maintenance and repairs. Stephane added: “We chose this instrument because the monochromator system allows us to work with any wavelength we need without considering whether or not we have the correct filters.

Also, its resistance to decontamination products like hydrogen peroxide is essential for our work. It is simple to use, a convenient size and, like all Tecan instruments, is known to be a robust device.”

The excellent performance and organizational methods of the P4 laboratory have been recognized by the European authorities and it is now coordinating the European Research Infrastructure on Highly Pathogenic Agents (ERINHA) project, aiming to provide better response and protection for citizens in the event of an epidemic.

To find out more about Tecan’s Infinite 200 PRO plate reader, visit www.tecan.com/infinite200pro To find out more about the P4 Inserm-Jean Mérieux laboratory, please contact directeur.laboP4-lyon@inserm.fr

Roland Durner, General Manager for Tecan Australia

Leading the debate Tecan’s customers in the Australasian region want the same thing as all our customers; reliable, intuitive, state-of-the-art products, backed by applications know-how, technical expertise and flexible local support. However, in our region perhaps more than anywhere else in the world, this can be easier said than done, as some of our competitors have discovered.

Many laboratories in Australia and New Zealand, in universities, hospitals and government research institutions, are funded by public money. Safety and security are crucial aspects, and it is critical that instruments are used to full advantage and are always up and running, not sitting collecting dust.

Purchasing decisions are frequently made based on the availability and track record of a supplier’s maintenance and repair services.

Australia and New Zealand are massive exporters of agricultural products.

Tecan Australia supports the quality, product development and food safety departments of these customers, who are often situated in very remote locations. No matter how reliable an instrument, the option to have maintenance and support services available is a big consideration before making any capital outlay.

Providing the capacity to support highly sophisticated customers, servicing complex instruments – both Tecan branded and those produced through its partnering business – far away from the company headquarters is what really sets Tecan Australia apart. Tecan Australia combines reliable products with a flexible, local support organization and, in its role as Tecan’s newest subsidiary, it is well placed to support our customers, wherever they are and whatever they need, in our region.

How crucial is local support to you? Send your comments to talk@tecan.com

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EVENTS 2012 TECAN JOURNAL 1/2012

Meet Tecan at these events Americas SLAS Society for Laboratory Automation & Screening

San Diego, California

04 – 08 Feb 2012

MD&M Medical Design & Manufacturing West

Anaheim, California

13 – 16 Feb 2012

AAFS American Academy of Forensic Sciences 2012

Atlanta, Georgia

20 – 25 Feb 2012

Pittcon Conference & Expo 2012

Orlando, Florida

11 – 15 Mar 2012

ABRF Association of Biomolecular Resource Facilities 2012

Orlando, Florida

17 – 20 Mar 2012

24th Lorne Cancer Conference

Lorne, Australia

09 – 11 Feb 2012

Japanese Society for Bioscience, Biotechnology and Agrochemistry

Kyoto, Japan

22 – 25 Mar 2012

Asia and Pacific

CMEF China International Medical Equipment Fair Spring 2012

Shenzhen, China

17 – 20 Apr 2012

BIO tech 2012

Tokyo, Japan

25 – 27 Apr 2012

Brussels, Belgium

22 – 23 Mar 2012

Europe, Middle East and Africa Laborama 2012 Gen2Bio

Lorient, France

29 Mar 2012

ArabLab

Dubai, UAE

26 – 29 Mar 2012

Analytica

Munich, Germany

17 – 20 Apr 2012

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.

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 Austria +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 www.tecan.com Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Marc Wetli/Zürich www.wetli.com, Günter Bolzern/Zürich www.bolzern.net, Susanne Völlm/Zürich www.susannevoellm.ch Editor in Chief: Tecan Trading AG, Cornelia Kegele Project Lead: Tecan Trading AG, Cornelia Kegele/Tanja Sager 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 © 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