Tecan Journal Life Sciences
Clinical Diagnostics
Forensics
OEM
Business and Technical News
Edition 2 / 2009
Oceanic research to predict global climate change pages 18–19
Control at your fingertips – TouchTools Suite™ page 5
An automated answer for molecular pharmacology pages 12 – 15
Small volume DNA quantification with high throughput pages 20 – 21
CEO WELCOME TECAN JOURNAL 2/2009
Welcome to a new edition of the Tecan Journal... In these times of global economic hardship, it is more important than ever to have strong, reliable partners in industry on whom you can rely. Tecan is willing and more than able to fulfil this role. We continue to invest healthily in innovation, despite the financial crisis that is affecting other players in the life science market. Following a hugely successful year in 2007, figures show that Tecan’s net profit margin improved further in 2008, to 12.9 % from 12.6 % in 2007. We also have a long and successful history of involvement with several national and international funding initiatives, including the NIH in the US. Remember, our experience could be invaluable to you when you need to apply for programs and grants.
To reflect this current climate, this mid-year edition of the Tecan Journal shows many ways in which our solutions have saved our customers time and money, bringing the benefits of automation to a range of applications. We have details of how our liquid handling systems are helping to reduce industry costs by streamlining drug discovery and process development activities, and look at some exciting uses of HS Pro hybridization stations in the biopharmaceutical industry, including a program at MIT exploring how to prevent HIV infections developing into AIDS. An exciting story from Japan shows how Tecan’s technology has benefits in all walks of life, and there’s news too about how the new TouchTools Suite™ makes our automated systems even more accessible than before. Enjoy this issue, Thomas Bachmann, CEO
Welcome
2
CONTENTS TECAN JOURNAL 2/2009
pg 5
The development of TouchTools Suite™ is a perfect example of how Tecan personnel live and breathe automation! pg 8-9
2
CEO welcome
4
Higher throughput with CrysScreenTM
4
FE500proTM – pre-analytics made easy
5
Control at your fingertips with TouchTools SuiteTM
6
Automated cell culturing and analysis
6
LabAutomation 2009
7
Sharpening up our market intelligence for mutual benefit
8-9
Clever tip packaging to save on space and waste
10-11 Flexibility and efficiency four times over with Freedom EVO® 12-15 An automated answer for molecular pharmacology pg 7
pg 22-23
Automation aids nationwide surveillance system for virus outbreaks
Contents
16-17 ACCESSing yeast genomics 18-19 Oceanographic research to predict global climate change 20-21 Small volume DNA quantification with high throughput 22-23 Automation aids nationwide surveillance system for virus outbreaks 24-25 Biopharmaceuticals – manufacture in miniature 26-27 Bringing FISH home 27
Get your views across with T@lk to Tecan
28
Events 2009
3
4
PRODUCT NEWS TECAN JOURNAL 2/2009
Higher throughput with CrysScreen™ CrysScreen software provides crystallographers with an intuitive user interface to help manage complex protein crystallography protocols on Freedom EVO® workstations. The software guides users through automation of protein crystallography trials, including buffer creation, set-up of experimental conditions, running experiments, data management and results analysis. The latest release of the software offers higher throughput than ever before, with an enhanced worklist generator to allow parallel pipetting tasks on a single instrument, and straightforward control and review of project management using the powerful Project Tree. To ensure compatibility with your laboratory workflow, CrysScreen is closely integrated with Tecan’s Freedom EVOware® instrument control software, and has now been updated to guarantee full compatibility with Windows Vista® for simple control of hanging drop, sitting drop and microbatch under oil crystallization experiments. To find out more on Tecan’s CrysScreen, visit www.tecan.com/crysscreen Windows Vista is a registered trademark of Microsoft Corporation.
FE500pro™ – pre-analytics made easy Automation is fast becoming the method of choice for clinical laboratories facing demands for increased throughput and traceability. The FE500pro is Tecan’s front-end solution for automation of preanalytical processing, improving process security and safety, while reducing costs and allowing staff to concentrate on other tasks. The FE500pro offers walkaway processing, incorporating a wide range of functions onto a compact system, including: ̴ z _sY( v(=Fz v Fg` `$ sv(̟zgv F`= ̴ (` vF2 = Fg` ̴ YFu F$ Y( (Y (vF3 Fg` ̴ Yg $( ( Fg`
̴ $(̟ ssF`= ̴ z( g`$ v ( Y (YF`=
To find out more on Tecan’s FE500pro, visit www.tecan.com/fe500
̴ YFu g F`= ̴ zgv F`= g2 (z F` g ` Y (v v Vz The FE500pro workstation.
The FE500pro has an enclosed worktable to ensure biosafety and minimize risk of sample contamination, and the platform’s twin lane conveyor maximizes processing flexibility. The system uses automatic barcode and liquid level verification to guarantee security of patients’ samples during processing, and communicates directly with your LIMS to generate the correct number of secondary tubes for your workflow.
PRODUCT NEWS TECAN JOURNAL 2/2009
Control at your fingertips – TouchTools Suite™ The development of TouchTools Suite is a perfect example of how Tecan personnel live and breathe automation! One day in 2006, on a long drive through Europe, three Tecan specialists came up with a new way to make Tecan’s automated systems even more accessible, particularly for users who are not familiar with writing software scripts. Eskil Trollhagen, product manager for Freedom EVOware® and TouchTools Suite, Michael Laab, head of Tecan’s Software Competence Center in Mainz, and Beat Glauser, program manager and now senior project manager at REMP, began by discussing possible software innovations that would complement robotic instruments, and a small, unofficial feasibility study soon resulted in the first prototype touchscreen interface. The positive reaction from customers to this first prototype propelled the development of the TouchTools Suite, which was launched at LabAutomation 2009 earlier this year to great acclaim.
The whole concept behind TouchTools Suite is that of a simple touchscreen. The appealing user interface allows easy control of the pipetting platform with a number of functions. The Application Starter offers a selection of methods from a graphical list, guides the user through the start procedure and provides on-screen instructions for loading the worktable. Instant Pipetting™ is a unique, patent pending module for direct, real-time control of pipetting robots. Even without programming skills or a pre-defined script, the user can immediately start pipetting without help from an automation specialist. Pipetting actions can be repeated or saved, effectively providing a new way of programming the instrument. QuickStart presents buttons on the touchscreen that can be linked to specific scripts so that chosen applications can be started immediately, literally at the touch of a button.
Carroll Ray, our travel agent, trains Sally Spafford, Tecan’s purchasing agent, after only five minutes of experience with the TouchTools Suite.
The Common Notification System (CNS) allows the user to monitor an instrument’s status remotely, notifying them of unexpected events thereby increasing walkaway time and process security. CNS can now be accessed through iPod touch, iPhone or other mobile devices with a web browser. The new Freedom EVO® Remote application, available in the iPhone App Store, allows convenient monitoring of all the instruments in the network, which can be particularly useful if the instrument is in a clean room, sterile hood or cold room, where access is difficult and could risk contamination. The TouchTools Suite launch at LabAutomation 2009 saw crowds flocking to the Tecan stand to try Instant Pipetting for themselves. www.tecan.com/touchtoolssuite
iPod is a registered trademark, iPhone is a trademark and App Store is a service mark of Apple Inc. in the US and other countries. Microsoft, Windows and Windows Mobile are registered trademarks of Microsoft Corporation in the US and/or other countries. Wi-Fi is a registered trademark of the Wi-Fi Alliance.
5
6
CORPORATE NEWS/COMPONENTS TECAN JOURNAL 2/2009
Automated cell culturing and analysis Tecan and innovatis are working closely together under a new collaborative agreement to jointly market their robotic cell culture workstations and cell detection devices. The collaboration concentrates on Cellavista, innovatis’ new automated image-based platform technology for cellular analysis, which has bright field and fluorescent capabilities and can process microplates, microscope slides and robotfriendly cell culture flasks. Cellavista, in combination with Tecan’s flexible Freedom EVO® platform, allows automation of a wide range of cell-related processes including cell line development, cell proliferation, stem cell differentiation and various cell-based assays. “We have already had a long, successful partnership with Tecan, and have worked very closely with Tecan application specialists on several customer projects,” said Michael Grohmann, Chief Executive Officer at innovatis. “Most Cellavista customers ask about full process automation and have shown great interest in Tecan’s automated solutions. They readily recognize the advantages of having a liquid handling and cell culture system that complements the detection capabilities of the Cellavista.”
Matthew Robin, Senior Vice President and Head of Liquid Handling and Robotics at Tecan, added: “The Cellavista has excellent image quality, fast reading speeds and versatile image recognition software, perfectly complementing our automated handling systems. We already have fully operational installations at several sites, for applications in stem cell differentiation, cell quality control and cell line development. I am delighted to continue Tecan’s fruitful association with innovatis, helping us to offer our customers a powerful package for cell-based applications.”
The Freedom EVO workstation, combined with innovatis’ Cellavista automated image-based platform technology, allows full automation of a wide range of cell-related processes.
LabAutomation 2009 community and providing manufacturers with an opportunity to showcase their latest systems. Among the new Tecan products on display were the Cavro® Centris Pump and Cavro Omni Robot.
Tecan enjoyed another successful exhibition at the ALA’s LabAutomation conference in January 2009 in Palm Springs, California. This annual gathering brings together world leaders in laboratory automation, highlighting new developments in the global
Tecan Cavro OEM components offer instrument developers a comprehensive hardware and software infrastructure for liquid handling applications, for use throughout the design process to accelerate development of new systems. In a workshop to help instrument designers take full advantage of these modular solutions, Tecan Systems invited existing OEM customers – BioDot and Merit Automation – to share their experiences of using Tecan Cavro products. David Wold, a senior product
l to r: Sean Leu (Software Manager, Tecan Systems), Drew Miller (Merit LabAutomation), Tony Lemmo (Vice President R&D, Bio-Dot Inc.) and David Wold (Senior Product Manager, Tecan Systems), with the new Tecan Cavro products at LabAutomation 2009.
manager at Tecan Systems, explained: “The workshop gave automation designers an opportunity to see how Tecan Cavro modules can help get their systems to the market faster, with BioDot and Merit Automation offering alternative perspectives and highlighting the potential advantages of our products. Overall the show was a great success, and we hope to be back for another workshop next year.”
QUALITY TECAN JOURNAL 2/2009
Sharpening up our market intelligence for mutual benefit Tecan’s Regulatory Affairs (RA) team has invested in a new market intelligence infrastructure that will keep our product development teams, business units and worldwide sales organizations, as well as our customers, fully up-to-date with regulatory information as it changes.
The current regulatory situation in the World today
What is our target?
Medical products companies are striving to increase their baseline sales by meeting the demands of today’s fast-growing, competitive global medical device market. Success can be limited by many factors, including new or changed international import and export regulations and/or registration requirements.
We won’t be caught out by changes! Tecan’s RA professionals are vigilant and proactive. It is our responsibility to ensure that effective market intelligence is collected for all countries where Tecan is operating, now and potentially in the future. The market intelligence we receive through our new system is continually updated and, as we receive new information, we will perform gap analysis to identify what is necessary to adapt to changing requirements. We react immediately to any new requirements that might hinder Tecan’s ability to operate in some countries.
Are there more things to be considered? Yes, Tecan RA professionals will also improve the service for you on currently marketed or registered devices. In addition to the market surveillance tool, a dedicated worldwide device approval monitoring system is currently being introduced. This enables the consolidation of all existing registration information such as: ̴ product classification
It is our intention to always be one step ahead of our competitors, and to ensure sustainable compliance with all global market registrations and regulations as they relate to the standards with which we must comply. One of RA’s goals is to meet the demands of Tecan and our customers.
How do we realize that? RA’s senior management has been very supportive towards our goals and objectives, allowing us to increase our market surveillance activities and to establish a powerful state-of-the-art software database. The database provides regulatory intelligence dossiers organized on a countryby-country basis – currently including 63 countries – which include country-specific reports, forms, and other useful background information. Tecan RA professionals already have unlimited and immediate access to this information. This will keep our product development teams, business units and worldwide sales organizations, as well as our customers, fully up-to-date with regulatory information as it changes.
̴ z _FzzFg` $g _(` z ̴ (z (v F3 (z ̴ řƍŵ (v F3 (z ̴ v(=Fz v Fg` (v F3 (z `$ _ C _gv( country-specific information
What are the benefits? You will notice real improvements in the response to your regulatory enquiries; we will respond in a faster and more uniform way. In addition, Tecan and customer RA professionals around the world will be able to work more closely together by using the same information source. We believe that having good quality information is essential for your decisionmaking process, whether you’re releasing a Tecan product into a market for the first time or choosing to purchase a Tecan product. Our new system will help us to achieve our aim of providing you with the best regulatory market intelligence available.
7
8
SERVICES AND CONSUMABLES TECAN JOURNAL 2/2009
Clever tip packaging to save on space and waste Nested MCA™ 96 disposable tips from Tecan are now routinely used at Roche, Switzerland, not least due to the savings on space and waste compared to conventional disposable tips. In combination with the team’s Freedom EVO® systems, the new tips have also increased assay throughput. Scientists in the In Vitro Enzymology and Drug-Drug Interaction Group within the Drug Metabolism and Pharmacokinetics Department in Roche, based in Basel, Switzerland, use a total of five automated systems from Tecan, including two Freedom EVO 200 platforms, for their compound assays. Pascal Schenk, Biochemistry Assistant responsible for automation in the department, explained: “When we needed to upgrade our automated systems, which are also based on older Tecan platforms, it made sense to get a Freedom EVO system as the combination of speed, flexibility and accuracy it offered suited our needs. We run about 20 different experiments on our two Freedom EVO 200 systems and they are great to work with, as we have significantly improved efficiency with these systems. Many years ago now, our department carried out all the assays manually with very low throughput; a large assay which we now run in only four hours would have taken one year if performed manually, testing one compound at a time. Similarly, the work which it recently took a colleague a week to carry out using an old automated system, now only takes five hours on a Freedom EVO 200 system equipped with a 384-channel Te-MO™ module.” “My role is to take responsibility for programming scripts on Freedom EVOware®, and the training of any personnel in the department who work with our Tecan systems,” Pascal added. “I have also developed a collaborative relationship with Tecan, and regularly try out new ideas and products for them, and was one of the first to trial the new nested MCA 96 disposable tips. As we run very short pipetting intervals, we are using disposable tips to save time in our schedule, as we can then
The In Vitro Enzymology and Drug-Drug Interaction Group (l to r): Pascal Schenk, Stephen Fowler (head of laboratory) and Vittorio Bona.
leave out the washing steps. The MCA 96 disposable tips have been a great success for us, not least also because they are packaged in smaller boxes and require less storage space compared to conventional non-nested disposable tips. Space makes a real difference as this lab consumes about 900,000 disposable tips annually. Using nested disposable tips also means we generate far less waste compared to the previously available disposable tips and therefore save on waste disposal cost. Instead of 10 boxes each containing 96 disposable tips in one column on the Te-Stack™, we can have 40 inserts of 96 nested disposable tips in the same amount of room, giving us a four-fold increase in the number of tips stored in the same amount of space. The nested tips also allow shorter pipetting time intervals,
offering increased assay speed and assay organization flexibility. As Tecan has only changed the packaging of the tips, we still have the same accuracy and precision with these tips, with typical coefficients of variation (CV) of less than 2 % with volumes of 1 μl, which is really excellent.” Pascal continued: “Each of our two Freedom EVO 200 platforms has several modules. The first system is also used for transcellular transport assays, and includes a multitude of modules: a LiHa arm, a RoMa arm, a MultiChannel Arm™ 96 option, five TeShake™ orbital mixers with heating plate (37 °C), a Te-Stack module storing nested MCA 96 disposable tips, a CO2 incubator and a special tilting carrier. The second platform has both a Te-MO 384 and an MCA 96.
SERVICES AND CONSUMABLES TECAN JOURNAL 2/2009
It also has a RoMa arm, a microplate reader, and two incubators each taking six 384-well plates, three Te-Shake orbital incubator/ mixers and a Te-Stack with the MCA 96 module and again nested MCA 96 disposable tips. The Te-MO 384 module handles the primary in vitro drug-drug interaction screening assay which determines the IC50 values for inhibition of drug metabolizing enzymes in human liver microsomes. Serial dilution of the test compounds in the 384-well plates takes place using the MCA 96 head and disposable tips and the plates are then transferred to the Te-MO 384 module, which pipettes all the reagents, starting solutions and quenching
solutions for incubations. The special feature of the Te-MO 384 module is its precision (CV <3 % for volumes of 250 nl) which allows total incubation volumes of 50 μl to be used with a DMSO content of 0.5 %. In addition, the existence of three spatially distinct cooling positions for the human liver microsomes and the start reagent greatly help with assay organization.” “Together, both Freedom EVO systems allow us to test 150 different compounds in one day in our higher throughput assays, leaving time for us to run the other standard assays, as well as some special ones. This flexibility offered by the Tecan systems has been vitally
The Te-Stack alongside the Freedom EVO workstation stores nested disposable tips for the liquid handling applications.
important for our assays, and, increasingly, other groups in our department are asking us about our systems and the assays we can run on them. We hope to purchase another Tecan system in the near future.” “I am really pleased with the Tecan systems and very happy with the support and close collaboration we have had with Tecan Switzerland. Using Tecan systems and disposable tips to automate our assays has dramatically changed how we do our work,” concluded Pascal. www.tecan.com/consumables Scientific instrumentation. Not for use in human clinical or in vitro diagnostic procedures.
The tilting carrier allows complete removal of liquids from the wells.
9
10
APPLICATION BIOPHARMA TECAN JOURNAL 2/2009
Flexibility and efficiency four times over Innovative drug discovery company Siena Biotech, based in Italy, has chosen four highly tailored Freedom EVO® systems for different roles within the company and each one has been invaluable in increasing throughput and accuracy in testing compounds as potential therapeutic drugs.
The Screening and Technologies team (from l to r): Marco Rossi (Oncology), Chiara Ghiron (Head of Screening and Technologies), Federico Cappelli (Biomolecular Screening), Marta Gherardi (Compound Management and Analysis), Wolfgang Fecke (Head of Biomolecular Screening), Raffaella Brandi (Compound Management and Analysis) and Hendrick Bothmann (Biomolecular Screening).
Siena Biotech S.p.A. is a drug discovery company whose research efforts are focused on diseases of the Central Nervous System, both neurodegenerative and proliferative and including rare diseases, with the aim of discovering new drugs for therapeutic intervention. In order to support the drug discovery process from target identification to clinical studies, the Company applies the most advanced technologies available in the biomedical and biotechnology fields. Siena Biotech is owned by the Monte dei Paschi di Siena Foundation, which is one of the 10 largest foundations in the world. The Screening and Technologies Department at Siena Biotech is headed by Chiara Ghiron and is comprised of a Compound Management and Analysis unit, a Biomolecular Screening unit, a Metabolic Profiling unit and a centralized Cell Culture laboratory. Four Freedom EVO systems equipped with various modules are used for a range of applications in the department. Chiara explained: “Our collective aim is to increase our efficiency both in higher throughput screening of compound libraries, and later in quality screening and compound
profiling phases. Automation is helping us to keep up with requests for a range of assays, from biochemical to high content readout formats. In Biomolecular Screening, two Tecan systems prepare assay plates for single point and concentration response analysis, pipetting all solutions and reading the plates by measuring absorbance, fluorescence or luminescence; in Compound Management and Analysis, a further system prepares compound plates in different formats for screening; while in Cell Culture, applications for the fourth system include plate coatings, seeding cells in plates and manipulating trans-well plates for permeability assays.” Hendrick Bothmann, Senior Scientist in Biomolecular Screening, explained: “I had worked with Tecan systems before I came to Siena Biotech, and recommended that we approach Tecan when we were experiencing problems with an existing automated system. We
eventually replaced the old instrument with a Freedom EVO system and, from then onwards, the excellent service we are receiving is making all the difference. The high flexibility of the Tecan systems, including the ability to integrate other instruments is the first thing I like about them. Secondly, the open concept of the Tecan software gives more possibilities for integrating systems and setting up assays which require non-standard operations. One example of this is our scratch assay; this assay tracks the time taken for cultured cells to move over a space on the bottom of the well created by scratching the cell layer with a 50 μl pipette tip. This gives us important information about whether test compounds affect cell migration, for example for cells from glioblastoma multiforme, one of the most common and aggressive brain tumors. Performing this test manually was too timeconsuming with low reproducibility; an automated method was essential to achieve precise width and direction of the scratch, and validate this using imaging software. Working closely with Tecan, I designed a vector that works really well, allowing the pipette tips to touch the bottom of the wells and scratch over a defined distance.” Federico Cappelli, Research Technician in Biomolecular Screening, added: “We are still in the early stages of adapting some of the existing manual assays for our Freedom EVO system, and we are currently working on automating our Madin-Darby canine kidney (MDCK) cell line permeability
APPLICATION BIOPHARMA TECAN JOURNAL 2/2009
Freedom EVO platform with multichannel arm in Biomolecular Screening.
assays in collaboration with the Metabolic Profiling Unit. The flexibility of the system will allow us to reproduce the results currently obtained by a dedicated operator, increase the throughput and minimize interoperator variability.” Marta Gherardi, Research Technician in Compound Management and Analysis, continued: “After hearing about Hendrick’s and Federico’s experiences with the Tecan systems, we chose our own Freedom EVO system to improve throughput, customized to exactly meet our needs thanks to the Tecan Integration Group (TIG). We visited TIG in Switzerland and looked at all the possibilities for customization, choosing the ones that best suited us. The system was then built in Switzerland and integrated on-site with our automated compound storage system.” Raffaella Brandi, Scientist in Compound Management and Analysis, added: “When our customized system arrived, it only took one week of testing before it was ready for routine tasks, allowing plates to be retrieved from and returned to the storage system without manual intervention, and giving greater throughput and longer unattended run times.” Marta continued: “The ability to leave the Freedom EVO unattended while it is plating for three or four hours at a time, or even overnight, is very appealing. The Freedom EVO can potentially be left running for days as we have a sealer for our compound plates. Running the system to its full capacity is something we will consider in the future to accommodate increases in our compound screening workload.”
Hendrick concluded: “Tecan has offered us training when we needed it, tailoring it to our requirements, and the service is outstanding and very efficient.” Federico agreed: “Our interaction with the Tecan Service Engineers in Milan has been extremely important and beneficial. They have taught us many things we needed to know for our routine applications.” www.tecan.com/tig Scientific instrumentation. Not for use in human clinical or in vitro diagnostic procedures.
The Freedom EVO platform in Compound Management and Analysis prepares compound plates for screening.
11
12
TECAN INTEGRATION GROUP TECAN JOURNAL 2/2009
An automated answer for molecular pharmacology The Leibniz-Institut für Molekulare Pharmakologie is using Tecan’s Freedom EVO® workstations for systematic high throughput screening of bioactive small molecule libraries for potential research tools and targets for drug development. These systems form the hub of ChemBioNet’s Screening Unit, providing an interdisciplinary open access platform in support of academic research projects exploring biological function. an ideal environment for research projects aiming to discover biologically active substances.
The team at the Screening Unit (front row, l to r): Dr Martin Neuenschwander and Dr Simone Graeber; (back row, l to r): Dr Silke Redetzky, Andreas Oder, Christoph Erdmann, Carola Seyffarth, Dr Jens Peter von Kries and Chris Eckert.
The Leibniz-Institut für Molekulare Pharmakologie (FMP) in Berlin, Germany, is a major international contributor to the field of molecular pharmacology. The FMP’s research focuses on the structure, function and pharmacological interaction of proteins, compounds and small biomolecules to identify structures and molecular scaffolds of interest to both fundamental biology research and drug development. The Institute’s broad interdisciplinary expertise and state-of-the-art technology place it at the forefront of drug development, as well as maintaining close ties with many academic organizations conducting fundamental research.
The FMP also plays host to the Screening Unit of ChemBioNet, an initiative by biologists and chemists in academia to provide an interdisciplinary open access platform in support of research projects exploring systematic usage of small molecules in biological systems. This initiative aims to provide access to high throughput technologies, allowing identification of compounds useful for dosage dependent, temporal or localized interference with biological function. The unique combination of technology available at the FMP includes the open screening platform, a state-of-art NMR facility and a mass spectrometry laboratory, providing
The FMP’s Screening Unit also manages a variety of technology platforms for other ChemBioNet members, working closely with the Max Delbrück Center for Molecular Medicine (MDC), the Helmholz Institute for Infection Research, the University of Oslo and the Berlin Institute for Medical Systems Biology (BIMSB). The FMP’s advanced technology and unique position within the European research environment offer unprecedented opportunities for the study of chemical biology, providing a research hub to enable close collaboration between academic institutions. In addition, the Institute’s structure allows a very broad range of projects to be investigated, using the same technologies available to the pharmaceutical industry but without the limitations associated with drug development, as Dr Jens Peter von Kries, Head of the Screening Unit, explained: “The research projects conducted at the FMP cover a wide variety of pharmacologically and biologically relevant pathways, and it is not essential that these are associated with a known disease state. Not being restricted in this way is very important to the way the Institute works, and this broad approach often leads to a surprising level of insight into the mechanisms of disease, even if the relevance of the research is not immediately and obviously relevant. It is
TECAN INTEGRATION GROUP TECAN JOURNAL 2/2009
important for us not to try and compete with the pharmaceutical industry, which can easily lead to duplication of research, and the major focus of the FMP is on protein-protein interactions, which have not historically been of interest to pharmaceutical companies.” “The Screening Unit has a small team of staff performing compound library screens, and we work very closely with colleagues from other departments and institutions for both analysis of the data we generate, and to search global databases such as the World Drug Index for fragments of molecular scaffolds and sub-structures of interest. Our platform has a very high throughput, allowing us to screen an average of 20,000 compounds a day, and this capacity is partly thanks to the flexibility of our Freedom EVO workstations. Many of our projects use either high content screening with automated microscopes or genome-wide RNA interference (RNAi) studies, and we have a Freedom EVO system configured for each of these methods, and an additional platform for management of the ChemBioNet screening library. Each workstation has been collaboratively designed with the Tecan Integration Group (TIG) to create a robust system that is capable of very high throughput analysis, giving consistent results even with continuous use.”
A Freedom EVO workstation at FMP, Berlin.
13
14
TECAN INTEGRATION GROUP TECAN JOURNAL 2/2009
High content screening
Genome-wide RNAi
Compound management
“
The RNAi workstation has a very similar specification to the high content platform, without the automated microscope of course, and is used predominantly for studies of human and C. elegans cell lines in association with the MDC and BIMSB.
“
The ChemBioNet screening collection is shared with many academic institutions in Germany and across Europe, and a Freedom EVO workstation with a REMP SmallSize Store™ (SSS) is used for both management and screening of this collection, with newly synthesized or purified compounds being added all the time. This workstation uses both an MCA™ 96 multi-channel arm and a LiHa arm for liquid handling, as well as a range of REMP sample storage modules, including the SSS. This is a very flexible system, bringing the powerful technology of larger REMP cold stores, which have become the industry standard in many sectors, to research laboratories. Our system was used to alpha test integration of the SSS into the Freedom EVO platform, and the resulting Freedom EVO / REMP SSS Factory represents the ideal combination for screening applications, improving throughput, maximizing walkaway time and allowing full sample traceability, while protecting precious library samples from denaturation, contamination and decomposition.
Our high content screening platform uses an automated microscope system, designed specifically for live cell imaging. This is integrated into the Freedom EVO workstation, and the whole system operates within a sterile environment. This system also has two automated nitrogen incubators at the back of the worktable for storage of cell culture flasks, compound libraries and 384-well test plates. The system is equipped with both 384-channel TeMO™ and 8-channel LiHa pipetting arms, allowing complete automation of the screening process, including harvesting of cells from culture flasks and parallel pipetting of candidate compounds and controls for fluorescent imaging. The microscope software documents images and identifies objects, converting fluorescence intensities to multi-parameter tables for effective analysis of screening data. This enables measurement and documentation of cellular kinetics and events in 384-well format in just a few hours, with reaction volumes of just 20-40 μl.
”
The human genome RNAi library encompasses approximately 20,000 genes, and our close association with the MDC means that many of the RNAi projects are relevant to various cancers, particularly control of metastasis. By contrast, a majority of the work done with C. elegans cell lines is for systems biology studies, and we have a library of over 16,500 strains, covering around 87 % of C. elegans genes. For analysis of RNAi studies we are using a variety of optical methods, including both fluorescence and chemiluminescence techniques, and the Freedom EVO platform allows us to integrate several different reader technologies into the workstation.
”
“
”
TECAN INTEGRATION GROUP TECAN JOURNAL 2/2009
Screenshot of siRNA screening.
“As well as being able to integrate numerous robotics and detection modules, it is essential to have a straightforward and flexible software configuration. Tecan’s Freedom EVOware® delivers this, allowing management of the entire screening process by directly interacting with control software for storage devices, incubators and readers, and has many innovative and unique features to provide robust control of screening assays with full documentation of every step. Tecan’s software engineers have always provided us with excellent support for integration of new devices, and have also modified drivers as software configurations have been updated. This means that we are now able to use one software solution for complete control of screening, where previously two or three different packages would have been required. This simplifies sample tracking and documentation, making the system faster and easier to use.”
both factory and site acceptance tests to guarantee that the platform exactly matches our needs. Validation of the system is then very simple, and Tecan staff are very willing to support us with further development of the platforms as new technologies become available. The core expertise of our staff at the Screening Unit is predominantly in biology or chemistry, and so we are dependent on our hardware suppliers’ experience in biological process automation. Tecan takes a keen interest in its customers’ projects, and our staff are trained by Tecan to a high standard to ensure we are able to maximize the potential of both our platforms and software. We enjoy a very
“Each of our systems was carefully designed to meet our requirements, and the team at TIG have worked hard to make sure that the final configuration represents the best possible solution available. The approval procedure is central to this process, with
Tecan Integration Group.
good working relationship with Tecan’s automation engineers, receiving immediate expert support for all our projects. Tecan has an exceptional culture of customer service, and our work here at the FMP undoubtedly benefits from this close partnership.” To find out more on Tecan’s Freedom EVO workstations, visit www.tecan.com/freedomevo To find out more on Tecan Integration Group, visit www.tecan.com/tig Scientific instrumentation. Not for use in human clinical or in vitro diagnostic procedures.
15
16
LIQUID HANDLING AND DETECTION TECAN JOURNAL 2/2009
ACCESSing yeast genomics
Tecan systems are providing an integrated solution to researchers at Stanford Genome Technology Center for the study of yeast genomics. The laboratory’s ACCESS robot project is combining several Tecan technologies to develop a fully automated platform for phenotype analysis.
l to r: Ulrich Schlecht, Julia Oh and Michael Proctor.
Although the yeast genome is relatively small in comparison to higher eukaryotes, this ‘simple’ system is still comprised of nearly 6,000 genes spread across four chromosomes and its study is very relevant to the understanding of complex cell biology. The Genome Technology Center at Stanford University in California, USA, in collaboration with the University of Toronto, Canada, has a strong research interest in the functional genomics and chemogenomics of yeast, and is developing powerful tools to assist in the investigation of this model organism. The Center currently has several yeast research projects running, including establishing a growth phenotype for every gene in the genome, reciprocal hemizygosity studies (to characterize complex multigene phenotypes) and screening libraries of small molecules for compounds affecting yeast phenotype. Such ambitious projects require high throughput genetic techniques, and the Center is at the forefront of developing new genomic technologies including an automated cell, compound and environmental screening system (ACCESS) robot for haplo-insufficiency and overexpression profiling in yeast(1, 2), as well as high resolution phenotypic analysis of other single cell organisms. Dr Michael Proctor, a research and development scientist at the Stanford Genome Technology Center, explained their investigations: “Our laboratory’s work, in association with our collaborators Dr Robert St Onge from Stanford, and Dr Guri Giaever and Dr Corey Nislow from Toronto, is focused on establishing the functional genomics of S. cerevisiae, using a combination of chemogenomics and knockout mutants to establish the phenotype associated
with up or down regulating any given gene. Phenotypes are often quantitative and influenced by multiple genes, so it is important for the data we generate to be robust and reproducible, and this is where automation is a great advantage. Another advantage of automation is that it allows the development of experiments which would otherwise be too complex or errorprone to be run routinely.” “For a number of years we have been developing the ACCESS platform to eliminate many of the manual steps of plating, growing, screening and data analysis of yeast cultures in liquid medium. This has resulted in much higher resolution data and improved reliability, and we are now able to identify very small changes in the growth curves of our yeast cultures. Our latest system is based around a Freedom EVO® 150 liquid handling workstation, integrating incubator shakers, plate readers and pipetting stations to allow inoculation, re-plating and saving samples to a cold station. This system is designed to be extremely flexible, allowing multiple users to run different assays in parallel for maximum productivity. The Freedom EVO’s RoMa arm transports microplates between the various devices, and the LiHa arm performs inoculation steps as necessary.” “We have multiple temperature controlled plate shakers and a Safire2™ microplate reader arranged on our Freedom EVO platform, and are currently evaluating an Infinite® M1000 microplate reader for integration into the ACCESS system. This platform provides us with a high throughput capacity, enabling many cultures to be grown and monitored in parallel. We also
LIQUID HANDLING AND DETECTION TECAN JOURNAL 2/2009
have three non-liquid handling versions of ACCESS, for pre-screening and inhibitory concentration (IC) assays. These systems have up to seven Tecan microplate readers, which offer excellent condensation dissipation and temperature control characteristics. We run a combination of assays at any time, with 2,000 samples a day on average, although this is well below the capacity of the system using 384-well plates. When we first began using Tecan readers we used 24-well plates, but quickly progressed to 48-well plates. We now use a combination of 48- and 96-well plates for most of our screening work, although we have also achieved a high level of success with 384well plates. One of the greatest benefits of Tecan’s microplate readers is that they are not restricted to one plate format. You can use any plate type, and there is the option of scanning the plate and defining the parameters automatically using pre-set or customized protocols stored in a database, giving almost infinite flexibility.” “Operation of the ACCESS platform is controlled by LabVIEW™ software which has been developed in-house. This coordinates with our LIMS system, also developed in-house, and with the workstation’s Freedom EVOware® and the microplate reader software. It also schedules tasking of the instruments, and coordinates analysis of data. This approach simplifies complex pipetting operations for the user, and offers us the facility for rapid sequential measurements of optical density, fluorescence and luminescence as required. The software is also capable of automatically determining several growth parameters, autonomously governing inoculation, addition of screening compounds and
storage of samples. The LIMS is capable of large scale analysis and coordination of data and assays. This enables the ACCESS platform to run many multi-step, multi-plate assays with just a few barcode scans of the instruments, source and growth plates. ” “We have now been using Tecan instruments for over nine years, and over this time we have expanded our set-up to include more and more Tecan equipment, enabling us to very successfully increase the magnitude of our project. In total we have close to thirty Tecan instruments – spread over three laboratories in the USA, Canada and Germany – and these have been very reliable workhorses, shaking and reading every 15 minutes, 24 hours a day and running almost constantly as the ACCESS platform has developed over several years.”
The Freedom EVO platform at Stanford University.
Acknowledgements Dr Proctor would like to acknowledge the contributions of Malene Urbanus at the University of Toronto, Dr Robert St Onge and colleagues at the SGTC, and Mark Torresan at Tecan USA for their exceptional and productive collaboration. 1. Giaever et al (2004). Chemogenomic profiling: identifying the functional interactions of small molecules in yeast. Proc Natl Acad Sci USA, 101 (3), 793-8. 2. Hoon et al (2008). An integrated platform of genomic assays reveals small-molecule bioactivities. Nat Chem Biol, 4 (8), 498-506. LabVIEW is a trademark of National Instruments Corporation Scientific instrumentation. Not for use in human clinical or in vitro diagnostic procedures.
17
18
COMPONENTS TECAN JOURNAL 2/2009
Oceanographic research to predict global climate change The Japan Agency for MarineEarth Science and Technology (JAMSTEC) is involved in a wide range of basic marinerelated research, including observation and projection of the earth’s environment, inner earth dynamics research and study of marine life in extreme environments. To support its environmental monitoring, JAMSTEC has developed a miniaturized observation device to measure carbon dioxide (CO2) in seawater, relying on Tecan’s Cavro® XCalibur Pump for accurate automated pipetting.
Dr Yoshiyuki Nakano with the drifting buoy, alongside JAMSTEC’s TRITON buoy, a larger, anchored observation device.
JAMSTEC’s main objective is to collect and disseminate information that helps to understand changes in the earth’s environment, upon which the oceans have a big influence. Data are being collected by various methods and made publicly available for use in research and industry, as well as to help the planning and implementation of international environmental monitoring programs. JAMSTEC also develops basic technology for oceanic research, including data processing technology. For environmental monitoring, JAMSTEC uses research ships and buoys to measure seawater variables such as temperature, salinity and chemistry. Since 2005, funded by the Ministry of Education, Culture, Sports, Science and Technology, scientists at JAMSTEC’s Mutsu Institute for Oceanography (MIO) in Aomori, Japan, have been developing a drifting buoy device for automatic measurement of CO2
in seawater. “It is almost certain that CO2 released by human activity is the cause of global warming,” explained Dr Yoshiyuki Nakano, Research Scientist at MIO. “It is necessary to continue monitoring CO2 concentrations in order to predict future climate change. Although atmospheric CO2 is being measured in various locations, there are very few observations being made at sea – which represents 70 % of the earth’s surface – mainly because of difficult access, and because securing a source of electricity for the measuring instruments also presents problems. In situ seawater CO2 sensors are being developed in several countries, but all have issues with size or cost, and none are in widespread use.” “An important design goal was to make the CO2 sensor small and light, so it was necessary to have a compact pump. The Cavro XCalibur Pump was chosen for this reason, because the pump also offered high
COMPONENTS TECAN JOURNAL 2/2009
Oceanographic research vessel MIRAI (Japanese for ‘future’), which is being used for oceanographic surveys. One of its missions is to deploy observation buoys.
accuracy, was capable of maintenance-free operation for long periods, and a broad volume selection range.” Dr Nakano continued: “Our buoy weighs about 15 kg, light enough to be lifted by one person. It makes a measurement once a week and the results are sent to the laboratory via satellite. The device calculates the CO2 concentration in seawater, using the acidic nature of CO2 dissolved in water. Within the buoy, the CO2 in seawater passes through a gaspermeable membrane into a sample cell containing a pH indicator solution, which is analyzed by spectrophotometry. To help miniaturization, the capacity of this cell is less than 1 ml, and all connecting parts and tubes are very thin. Pure water and concentrated indicator need to be freshly mixed for each measurement because diluted indicator is quickly bleached by UV rays from the sun. Appropriate amounts of indicator and water are accurately aspirated using two Cavro XCalibur Pumps and mixed, before being put into the sample cell. The volumes of indicator and water used are critical; without sufficient accuracy, the final concentration of the pH indicator can be too concentrated or too dilute for reliable results, but the Cavro XCalibur Pump allows us to achieve exactly the concentration we aim for. The pump’s broad selectable volume range is important because the concentration of indicator to use has been determined empirically, and the pump
allows us to choose the exact concentration we want.” “In May 2008, we placed our first drifting buoy in Canada’s Sea of Labrador and, in January, the second in the Antarctic Sea,” said Dr Nakano. “We have just deployed another three in March 2009, this time in the western north Pacific Ocean. Once we have enough data, the information will be used for various research; JAMSTEC has its own program of computer-aided simulation research for future climate
prediction, but the data will also be made generally available worldwide to be used by other scientists too. Our work has only just started so the data are still of limited practical use but, in the future, I expect that it will be used by institutes like the Intergovernmental Panel on Climate Change (IPCC), an organization that has received the 2007 Nobel Peace Prize for its work towards securing neutral, objective information related to climate change,” concluded Dr Nakano.
TRITON buoy in situ.
19
20
READERS TECAN JOURNAL 2/2009
Small volume DNA quantification with high throughput Scientists from Affymetrix, Inc., a world-leading supplier of microarray equipment and assays, evaluated the Tecan Infinite® 200 NanoQuant multimode microplate reader and two other spectrophotometers for DNA concentration analysis in a new cytogenetic assay they have developed. The researchers showed that the Tecan instrument gave the best results, with ease-of-use, high throughput and a low coefficient of variation.
Diane Flaucher, senior research associate in assay development, preparing samples to be analyzed by the Infinite 200 NanoQuant.
The Product Development Organization at Affymetrix, in Santa Clara, California, USA, develops biological and molecular biological assays and solutions mainly for genomics applications used by commercial customers in industry and academia. Dr Jim Collins, Director of Assay Development Molecular Diagnostic, explained: “We have recently developed a new generation general cytogenetic assay, which requires the quantification of small volumes of DNA on a spectrophotometer.” Jim and his colleagues evaluated three spectrophotometers from different companies during the development of the new assay, including the Infinite 200 NanoQuant from Tecan.
“I had previously worked and collaborated with Tecan, and was very excited to see that Tecan had developed an instrument for low volume DNA concentration analysis, with a higher throughput than anything else available for quantification of low volumes. To discover an instrument that could process sixteen samples at once thrilled me. I have always been highly impressed with the quality of instrumentation from Tecan in terms of its robustness and reliability, and hearing about this new instrument fired my enthusiasm to evaluate it in our laboratory; I felt it could be highly suitable particularly for the new assay that we have been developing. We have a group of nearly ten
people working on this particular project, and the Infinite 200 NanoQuant is in regular daily use during assay development.” Vicky Huynh, Jim’s colleague who carried out the evaluation, explained: “I found the Tecan reader to be a fantastic addition to our laboratory because it is quick and the software is very easy to use, with automatic display of results and a summary at the end of the measurements. The NanoQuant Plate™ is compatible with multichannel pipettes for high throughput, and quantification of whole microplates is really quick, taking only a few minutes. The easy-to-clean plate helps to eliminate cross-
READERS TECAN JOURNAL 2/2009
contamination and there is no need to dilute samples. It is ideal for both high and low throughput, processing batches of sixteen samples in just one minute. I found that the instrument has a high reproducibility, with a coefficient of variation of less than 5 %.” Jim added: “The Tecan reader has worked really well for the assay, and is one of three recommended spectrophotometers included in our user guide accompanying the new assay. When we suggest instruments to our customers, our focus is to give them flexibility and provide them with reliable options. We want them to be able to use instrumentation that they already have in their laboratories where possible, but also recommend instruments when they need to buy specifically for the purpose to make the performance of the assay as good as it can be in their hands. The assay requires measurement of DNA concentrations on a spectrophotometer, and an increasing number of laboratories are beginning to move to instruments like the Infinite 200 NanoQuant where they can quantify DNA or RNA from very small volumes. This saves time and conserves samples, as dilutions are not required.” “In our experience, the Infinite 200 NanoQuant has advantages over the other instruments we tested, primarily the increased throughput and the features of the user interface. The Tecan instrument is very simple to set up runs; it measures the samples very quickly and the user interface gives an excellent dynamic graphic display that shows how the samples are progressing in real time. All these features make the instrument really easy to use and that is why we were very keen to provide it as an option to our customer base,” Jim concluded. To find out more, visit www.tecan.com/ Infinite200nanoquant
21
22
LIQUID HANDLING TECAN JOURNAL 2/2009
Automation aids nationwide surveillance system for virus outbreaks The Korea Center for Disease Control and Prevention (KCDC) in Seoul, Korea, conducts research to identify the etiology of communicable diseases and to find effective control, prevention, diagnosis and treatment for them. In its efforts to build a more advanced and specialized surveillance system, KCDC has chosen Tecan’s Freedom EVO® workstations to standardize automated protocols for the efficient detection of enteric viruses. Dr Doo-Sung Cheon of KCDC.
norovirus infections using recombinant virus-like particles, and for smallpox using live attenuated vaccinia virus, in a biodefense study.
The Korea Center for Disease Control and Prevention in Seoul, Korea.
KCDC is a leading institute for the control and prevention of infectious diseases in Korea and consists of six centers and 33 divisions, and is building close partnerships with other governmental agencies, international organizations, research institutes, private health corporations and academia, with the primary goal of protecting the health of the public. Within KCDC, the Division of Enteric and Hepatitis Viruses is responsible for the management of nationwide
surveillance systems for enteric virus infections, such as enteroviruses and noroviruses. Laboratory scientists perform molecular epidemiological research, and develop rapid diagnostic methods using various techniques such as ELISA-based immunoassays, immunochromatography and real-time reverse transcription PCR (RT-PCR). The division is also involved in basic research on the pathogenesis of enterovirus-induced myocarditis, and the development of vaccine candidates for
Dr Doo-Sung Cheon, senior researcher in the Division of Enteric and Hepatitis Viruses, explained: “Our main task is to organize and perform nationwide surveillance for enterovirus and viral gastroenteritis, collaborating with sentinel hospitals and 17 regional laboratories that belong to local institutes of Health and Environment. To perform this effectively, we wanted to improve our diagnostic methods by enhancing sensitivity in the detection of viral genomes in clinical specimens. We also needed to develop robust, unified methods that yield reliable results, irrespective of the regional laboratory’s environment such as the level of skill and experience of the staff, or the equipment available to them.” In 2006, the laboratory automated its viral nucleic acid purification protocol based on silica-coated magnetic beads, using a Tecan Freedom EVO platform combined with the Te-MagS™ module. This system offered a standardized procedure from nucleic acid extraction to the detection of the amplified viral genome, with reduced process complexity to minimize human error and false positive results caused by carryover.
LIQUID HANDLING TECAN JOURNAL 2/2009
Dr Doo-Sung Cheon and his colleagues in the Division of Enteric and Hepatitis Viruses at KCDC.
Hepatitis A (HAV), caused by an RNA virus found in bodily fluids and characterized by jaundice, weakness, and fever.
The protocol has been optimized to yield sufficient viral nucleic acid to be used as RTPCR templates, and can be adjusted using Tecan’s integrated software. To complement the automated method, the researchers developed a ready-to-use RT-PCR premix kit for the specific and sensitive detection of enteroviruses and noroviruses, and the Freedom EVO is also being used to formulate this mix.
by a norovirus contamination, we would be capable of processing about 150 samples per day with the automated process. A collaborating laboratory located in Seoul uses a Freedom EVO equipped with eight probes, and that system is able to process 96 samples per run within three hours, allowing the processing of about 300 specimens per day.”
Dr Cheon explained: “Before automation, we extracted nucleic acid manually using TRI Reagent® or column-based methods. Although we could process more samples using TRI Reagent, it is tremendously labor intensive and often led to false results due to experimental error. The automated system is both cost- and labor-effective compared to column-based extraction methods. The automation, together with our own reagent mixture, yields better RNA purity and concentration, and requires less investigator intervention; the eluates are automatically transferred to PCR tubes after nucleic acid extraction, and the researcher only needs to move the PCR tubes from the Freedom EVO to the thermal cycler.” “The Freedom EVO in our laboratory is typically used five times per week and 48 samples per day, taking 2.5 to 3 hours to process 48 specimens in one run so, if we were to encounter a large outbreak caused
The Division of Enteric and Hepatitis Viruses subsequently developed an automated system for the detection of norovirus genome using the same, Freedom EVO-based set-up. As a critical part of its project to establish an early detecting and alerting system for food-borne norovirus-induced outbreaks, the laboratory supplied all of the 17 collaborating regional laboratories with the Freedom EVO-based systems during 2007 and 2008. “Because our surveillance program relies on the network with the regional laboratories, the standardized protocol, made possible by automation, is very important to maintain the consistent quality of results. The Tecan workstation achieves reliable and consistent yields of RNA, giving assurance of quality for our laboratory data and the surveillance system overall, and it is now a critically important device for the work we do,” Dr Cheon concluded. TRI Reagent is a registered trademark of Molecular Research Center, Inc.
The Freedom EVO platform at KCDC.
23
24
APPLICATION BIOPHARMA TECAN JOURNAL 2/2009
Biopharmaceuticals – manufacture in miniature The Biomolecular Separation Engineering group at the Karlsruhe Institute of Technology, Germany, is using the Freedom EVO® liquid handling workstation, equipped with the new Te-Chrom™ module and Atoll’s 96-array MediaScout® RoboColumn system, to create a fully automated chromatography platform for biopharmaceutical process development.
The Biomolecular Separation Engineering group studies all aspects of downstream processing in the biopharmaceutical industry, focusing on rapid and optimized process development for industrial scaleup. Process development aimed at large scale production is generally highly timeconsuming and expensive due to the large number of potential variables and unit operations involved. Downstream processing thus accounts currently for over 85 % of the total production costs. To tackle this problem, the group has developed high throughput techniques to mimic all relevant unit operations used in the biopharmaceutical industry. These scaled-down models allow rapid and comprehensive automated characterization of proposed processes prior to pilot scale realization. The Te-Chrom module.
Professor Jürgen Hubbuch explained the team’s research strategy: “Originally, the group was involved in traditional experiencebased sequential process development; however, it soon became clear that time pressures of the biopharmaceutical industry made it very difficult to fully optimize and understand processes prior to scale-up. Around five years ago we changed our approach, switching to automation to help us learn as much as possible about processes before process synthesis and scale-up, aiming to significantly enhance process performance and reduce production costs. When choosing an automated platform, one of our major considerations
The Biomolecular Separation Engineering group (front row, l to r): Katrin Treier, Benjamin Maiser, Katharina Lang, Carolin Richter; (middle row, l to r): Anna Siudak, Stefan Oelmeier, Patrick Diederich; (back row, l to r): Jörg Kittelmann, Prof. Jürgen Hubbuch.
was that we wanted to be able to develop several new hardware components in-house. Therefore, we needed a flexible system that allows sufficient space to incorporate these modules and is compatible with the OEM robotics. We assessed all the automated liquid handling systems available, and found that Tecan’s Freedom EVO workstations offer unrivalled flexibility and a very high level of space on the deck of the instrument.” The group is using Freedom EVO workstations in conjunction with Atoll’s 96-array MediaScout RoboColumns, enabling fully automated parallel chromatography of up to eight columns. These columns are available prepacked with virtually all commercially available process separation resins, and column flow is controlled by the liquid handling arms via pressure tight inlets at the top of each column. Prof. Hubbuch continued: “As chromatographic separations are the heart of nearly all biopharmaceutical purification processes, one of the main tasks performed on our Freedom EVO workstations is assessing chromatographic separations of new biopharmaceuticals. One of the major advantages of the Freedom EVO platforms for this application is that liquid dispensing is hydraulically, rather than pneumatically, controlled. This is very important for chromatography, as pressure variations caused by compression of air in
APPLICATION BIOPHARMA TECAN JOURNAL 2/2009
0,14
A
A280 total (EXP) A280 RBN (Peak deconv.) A280 CTC (Peak deconv.) A280 LYS (Peak deconv.) A280 Sum signal
0,25
0,12
0,12
0,10
0,10
0,08
0,08
AU
AU
0,20
0,14
B
0,15
C
AU
0,30
0,06
0,06
0,04
0,04
0,05
0,02
0,02
0,00 0,0
0,00 0,0
0,10
1,0
2,0
3,0 4,0 Elution volume [ml]
5,0
6,0
7,0
1,0
2,0
3,0 4,0 Elution volume [ml]
5,0
6,0
7,0
0,00 0,0
1,0
2,0
3,0 4,0 Elution volume [ml]
5,0
6,0
7,0
Optimization (A-C) of a chromatographic separation of ribonuclease A, cytochrome C and lysozyme on the cation exchanger SP Sepharose FF. The separation was performed on a Tecan EVO Freedom 200 with Te-Chrom option and Atoll columns. (Blue diamonds: Experimental data from the Tecan EVO Freedom; blue line: fitted UV 280 signal of total protein content; red lines: de-convoluted signal of the individual protein species to be separated.)
the head space would affect flow rates through the columns. The combined Tecan / Atoll solution allows us to generate a large amount of data within a short time frame. The automation further allows us, for the first time, to exploit Design of Experiment (DoE) strategies combining short experimental time and low material consumption. This not only cuts the development time for process optimization significantly, but also gives a much better understanding of the process.” ”We now have multiple Freedom EVO workstations, and with each successive instrument we have revised our platform design to use the latest generation of Tecan modules. We have also developed several new ideas, including the Te-Chrom module to securely position the base of the RoboColumn plates during chromatography. Our systems have been optimized for high throughput analysis, using Te-Stack™ modules to increase walkaway time and enable fraction collection during elution.
Our latest system has a Te-Stack with a TeChrom, LiHa and MCA™ 96 pipetting arms, a RoMa arm to maneuver plates, and both a centrifuge and Infinite® 200 microplate reader below the deck. This set-up offers a high level of flexibility and maximizes deck space for our in-house hardware, and this is particularly important for another avenue of research we are exploring – miniaturization of complete industrial processes. By creating miniaturized versions of industrial components in a modular configuration, we are able to scale down the entire process to run on the Freedom EVO workstation. This enables us to have much better insight into how changing a variable, or fluctuation in a parameter, will affect production.”
EVO system in conjunction with Tecan here at the University of Karlsruhe. Automation is now essential to our research, and the flexibility of the Freedom EVO platform has enabled us to reduce process development time drastically. The significance of our work is highlighted by the fact that some of the largest biopharmaceutical companies are following this approach, using Freedom EVO workstations for downstream process development.” You can read more about the Biomolecular Separation Engineering group at: http://mab.ciw.uni-karlsruhe.de/ For more information on the automated
“Since acquiring our first Freedom EVO workstation we have worked very closely with Tecan, and later this year we will be taking delivery of our third Freedom EVO system. This will give us some extra flexibility, but will also be used for training. We are planning on running courses on how to use the Freedom
chromatography platform, visit www.tecan.com/proteinchromatography www.gelifesciences.com/bioprocess www.tecan.com/parallel and www.atoll-bio.com. MediaScout is a registered trademark of ATOLL GmbH
25
26
MICROARRAY PRODUCTS TECAN JOURNAL 2/2009
Bringing FISH home
Researchers at Medarex in California, USA, have semi-automated their hybridization process for FISH analyses with the HS 400™ Pro Hybridization Station from Tecan, allowing in-house screening of all their transgenic mouse strains.
Medarex, a biopharmaceutical company in California and New Jersey, USA, specializes in the discovery, development and potential commercialization of human antibodybased therapeutics for the treatment of life-threatening or debilitating diseases such as cancer, inflammatory, autoimmune and infectious diseases. The challenge faced by researchers is to convert the many thousands of potential disease-causing proteins – revealed by mapping of the human genome – into potential targets for new treatments. For many targets monoclonal antibodies may be the answer, and central to the research at Medarex is the production of high-affinity human monoclonal antibodies derived from transgenic mice. These strains of mice,
distinguished by the particular antibodies they generate, need to be characterized. Immunogeneticists Maria Galou-Lameyer and Poonam Sharma, members of Medarex’s animal biology laboratory, explained the role of fluorescence in situ hybridization (FISH) in initial strain characterization. “An important step in the characterization of our transgenic mice is using FISH, a cytogenetic technique, to visualize the location of specific nucleic acid sequences on metaphase chromosomes prepared from individual cultured cells. A fluorescentlylabeled probe, complementary to the target sequence, hybridizes to its counterpart within the chromosome, and the resulting fluorescence is detected using a microscope.
(from l to r): Maria Galou-Lameyer and Poonam Sharma with the HS 400 Pro Hybridization Station.
Probe of interest
Metaphase chromosome spread of a transgenic mouse. FISH with the probe of interest reveals the transgene insertion in green, while genomic DNA fluoresces blue with DAPI. The insertions are also visible in interphase nuclei.
TALK TO TECAN TECAN JOURNAL 2/2009
From FISH analysis of metaphase cells, we can establish the location and approximate copy number of the transgene in the mouse’s genome. The location of transgene insertion in the mouse genome can be critical to the level of expression, ultimately affecting both the quality and quantity of human antibodies produced in the engineered mice. The ability to raise high affinity antibodies to a wide range of therapeutic targets can be challenging, as not all targets have the same degree of immunogenicity. It is hoped that FISH techniques can help us to understand the relationship between transgene integration patterns in our mice, and their ability to mount a robust antibody response. FISH analysis also helps us to track the integration status of mice harboring multiple but unique transgenes. All of this work is ultimately about making better mice, and hence better therapeutic antibodies.” Dr Sharma continued: “Our FISH analysis begins with fibroblast cultures prepared from tail tips of very young mice. Metaphase preparations of these cultured cells are spread onto slides, then processed in our HS 400 Pro Hybridization Station. We chose this system because we have had plenty of good experience with systems from Tecan, so were very confident that this hybridization station would be reliable and deliver excellent consistency from experiment to experiment. The HS 400 Pro Hybridization Station automates several steps including pre-hybridization, addition of biotin-labeled probe, hybridization, post-hybridization washing and the addition of NeutrAvidin conjugate for visualization of the biotin-labeled probe. Four slides are processed per run, with a total run time of 21 hours, and we currently use the hybridization station three or four times a week. We need to process between 40 and 50 slides per mouse strain, giving a high workload during the characterization stage, however, no further screening is required once we have obtained good metaphase spreads to establish the integration pattern of our transgenes.” “The HS 400 Pro handles our current throughput needs very efficiently. It is an excellent workflowbased system, and has enabled us to start implementing in-house FISH analyses. This set-up will allow us to look back at specific transgenic strains, some of which were created several years ago, as well as newer uncharacterized strains. FISH analysis adds valuable information to the overall picture of the mouse genotype, which ultimately informs us about its phenotype. Having the HS 400 Pro has allowed us to work to our own schedule instead of being forced to rely on external contractors, where we only have limited control over throughput and experimental variables. Overall, it’s a very useful tool.”
Leading the debate Leading the debate in this issue of the Tecan Journal is Ralph Beneke, marketing product manager at Tecan for five years. In this auspicious year we celebrate the bicentenary of Charles Darwin’s birth and the 150th anniversary of the publication of his seminal work ‘On the Origin of Species’. What better a time to reflect on how the study of heredity and genetics has itself evolved, and what an impact this is having on society. Research depends on outstandingly innovative technologies making new discoveries everyday and, with applications encompassing clinical diagnostic and therapeutics, food testing, environmental monitoring and forensics, society benefits from genomics in so many diverse ways and its influence is closer to home than many realize. Genomics is moving forward so quickly that, for example, the latest generation sequencing technology is measuring at the level of single molecules – a truly amazing breakthrough. Technologies like high density microarrays are adding value to high throughput sequencing by their capability for fast front-end sequence capturing. At the same time, all genomic microarrays are capitalizing on new insights gained from ultra-high throughput sequencing providing better sequence and assay quality. The technology may change but it brings with it new challenges. How do we analyze the vast amounts of data this latest technology creates? How do we separate the noise from what is truly meaningful? And how can we rely on what we’re looking at? New bioinformatics tools and hardware are constantly being developed to deal with these new issues, but they are still part of the bigger picture that cannot be ignored. One remaining constant is that of good sample quality – however good the technology is, if you put garbage in, you get garbage out. Following along this theme, this year’s Tecan Symposium will focus on applied genomics and will present an ideal opportunity for researchers in these fields to discuss the future of this discipline.
Email talk@tecan.com to tell us what you think about this or another life science topic of your choice.
27
28
EVENTS 2009 TECAN JOURNAL 2/2009
Meet Tecan at these events in the coming months Americas ASM General Meeting
Philadelphia, PA
17-21 May 2009
AACC Annual Meeting 2009
Chicago, IL
19-23 Jul 2009
IIR Biorepositories Conference
Philadelphia, PA
21-23 Sep 2009
International Equipment Exhibition & Technical Seminar Of Forensic Science, Narcotics Control and Counter-Terrorism
China
20-22 May 2009
National Blood Collection and Supply of Laboratory Quality Management Conference
China
24-27 May 2009
8th International Bio Forum & Bio Expo Japan
Tokyo, Japan
01-03 Jul 2009
Australasian Association of Clinical Biochemists 47th Annual Scientific Conference (AACB) 2009
Brisbane, Queensland, Australia
14-17 September
8. Jahrestagung der Arbeitsgruppe Chipdiagnostik
Tutzing, Germany
14-15 May 2009
Advances in Microarray Technology AMT 2009
Stockholm, Sweden
19-20 May 2009
41st European Human Genetics Conference
Vienna, Austria
23-26 May 2009
Asia and Pacific
Europe, Middle East and Africa
ChemBio Finland 09
Helsinki, Finland
27-29 May 2009
21st Meeting of the European Society for Animal Cell Technology – ESACT
Dublin, Ireland
07-10 Jun 2009
Euromedlab 2009
Innsbruck, Austria
07-11 Jun 2009
ELRIG/SBS/BPS Drug Discovery 2009
Liverpool, UK
07-08 Sep 2009
11.Schweizerisches Symposium der Transfusionsmedizin BSD SRK/SVTM
Lugano, Switzerland
24-25 Sep 2009
BIOTECHNICA
Hannover, Germany
6-8 Oct 2009
MipTec
Basel, Switzerland
13-15 Oct 2009
Medica
Düsseldorf, Germany
18-21 Nov 2009
Biotech Forum and Scanlab
Stockholm, Sweden
25-27 Nov 2009
Tecan. For all your lab automation needs.
Liquid Handling & Robotics | Detection | Sample Management | Components | Services & Consumables
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 10 586 95 936 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 Portugal +351 21 000 82 16 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 ROW +43 62 46 89 33 REMP Switzerland +41 31 770 70 70 Japan +81 44 542 70 21 USA +1 508 429 2200
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/Aline Weiss 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 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.
www.tecan.com 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 CrysScreen, FE500pro, HS 400 Pro, Instant Pipetting, MCA, MultiChannel Arm, NanoQuant Plate, Safire2, Te-Chrom, Te-MagS, Te-MO, Te-Shake, Te-Stack and TouchTools Suite are trademarks and Cavro, Freedom EVO, Freedom EVOware and Infinite are registered trademarks of Tecan Group Ltd, Männedorf, Switzerland. Tecan is in major countries a registered trademark of Tecan Group Ltd., Männedorf, Switzerland. REMP Small-Size Store is a trademark of REMP AG, Oberdiessbach, Switzerland. © 2009 Tecan Trading AG, Switzerland, all rights reserved. To register for Tecan Journal please go to www.tecan.com/journal