Tecan Journal
Edition 2 / 2008
New 50 μl disposable tip for liquid handling platforms page 4
Developing one-step allergy testing panels page 14-15
Automating protein production processes for drug discovery at Bayer Schering Pharma AG pages 22-23
Ina Kaiser, Project Manager and Bernd Maisenhölder, Head of Product Management Consumables, Tecan Switzerland
2
C E O WE LCOM E
Focusing on your needs… …from designing new modules and consumables for specific applications, to working with academics to design innovative systems that will advance research and discovery, we aim to smooth the pathway for scientific progress. Welcome to this latest edition of the Tecan Journal, which highlights some of our recent customer-focused developments. Our family of high quality, disposable tips has been completed with the addition of the 50 µl volume size, which is ideal for genomics and forensics applications. In addition, and our new MultiChannel Arm™ 384 has been enhanced with the gripper module for even more efficient plate processing. We are delighted to bring you reports of several collaborations with leading biopharmaceutical companies including Bayer Schering Pharma AG. Furthermore, we have an exciting new project in the pipeline with regenerative medicine scientists at the University of Zurich, resulting from our ongoing support for increased communication between academia and industry. Read on to find out more! As part of our dedication to streamlining our processes, we are updating the Tecan Journal distribution database. We would be grateful if you could take a moment to re-register your current details online (www.tecan.com/get-journal), to ensure that you can continue to enjoy each issue. We hope you enjoy this issue, Thomas Bachmann, CEO
and don’t forget, to make sure you receive future copies of the Tecan Journal you’ll need to re-register your details online at:
www.tecan.com/get-journal
Tecan Journal 2/2008
CO NTENTS CEO welcome
Thomas Bachmann, CEO, welcomes you to another issue of the Tecan Journal.
Te-Care™ service contracts for peace of mind
page 2
Tecan’s network provides globally consistent service.
New 50 μl disposable tip for liquid handling platforms
Updated webpages launched
A new tip size completes Tecan’s range of conductive disposable tips.
page 4
PowerScanner™ – Tecan’s new flagship microarray scanner A new product to add power to your microarray experiments.
page 4
Added functionality with the MultiChannel Arm™ 384 gripper option A cost-effective alternative to a robotic arm.
page 5
page 8
New ‘consumables’ and ‘about Tecan’ webpages.
Automated pH measurements and solubility screening of candidate compounds in pharmaceutical development
Freedom EVO® workstation automates early stages of drug development at AstraZeneca UK.
pages 18-19
page 8
Automated pre-analytical blood sample sorting Tecan’s Genesis FE500™ saves time at Sahlgrenska University Hospital.
page 9
Cost-effective PAMPA with Infinite® M200
Tecan and pION validate drug permeability screening assays on the Infinite 200.
True high throughput compound management for Galderma
Asia Pacific Sales Meeting in Japan
pages 20-21
page 10
Freedom EVO® platform manages a compound library for dermatological drug development.
Tecan Japan and Tecan Asia Pacific delegates meet in Tokyo.
page 11
The Freedom EVOlyzer® takes routine clinical diagnostics tasks off your hands New software guides fully automated ELISA.
page 5
Automating protein production processes for drug discovery at Bayer Schering Pharma AG Automated differentiation of stem cells with the Freedom EVO® and Millicell®-24 filter plates Millipore automates stem cell culture and differentiation.
pages 12-13 Sub microliter pipetting with Te-PS™ tips Accurate low-volume pipetting for protein crystallography.
page 6
Simple control of complex sample pipetting procedures The latest version software for easy sample processing.
page 7
Making advances in regenerative medicine
Tecan collaborates with tissue engineering researchers.
page 7
Developing one-step allergy testing panels using the HydroFlex™ system
VBC-Genomics automate microarray washing with the HydroFlex.
pages 14-15
Efficiency by automation – high throughput sample processing
Genesis FE500™ automates sample handling in limited laboratory space.
page 16
Two Freedom EVO® workstations automate high throughput cloning, expression and protein purification.
pages 22-23
Automated screening systems in plant biotechnology
Advanced Technologies Cambridge automates PCR screening assays.
pages 24-25
Ahead of the game for quality processes New, trend-setting organizational structure for quality management at Tecan.
pages 26-27
Events 2008
Meet Tecan at these events in 2008.
page 28
High throughput EIAs with Freedom EVO® Clinical Freedom EVO Clinical runs thousands of immunological assays every week.
page 17
Tecan Journal 2/2008
3
4
and don’t forget
P ROD U CT N EWS
New 50 μl disposable tip for liquid handling platforms
PowerScanner™ Tecan’s new flagship microarray scanner Who wouldn’t want the best possible microarray scanner in their lab? The much anticipated, compact high resolution microarray scanning system will soon become reality: Tecan’s PowerScanner™, a new generation, automated high resolution microarray scanner, is due for launch in August 2008. As the new flagship of Tecan’s ‘microarray suite’ range, the PowerScanner features an ozone-protected environment for your valuable microarrays, and offers state-ofthe-art resolution, sensitivity, accuracy and reliability for automated processing of up to 48 slides in one run.
50 μl tips pipetting into microplate
Our family of high quality, conductive disposable tips has now been completed with the introduction of the 50 μl tip size, complementing our existing size range of 10, 200 and 1,000 μl. These disposable tips have been custom-developed for our automated liquid handling workstations, enhancing performance and minimizing cross-contamination in a broad range of genomics, forensics and ELISA-based applications. The 50 μl tip’s precision and volume range (1.0 to 50 μl) is optimal for PCR set-up and DNA quantification and normalization, as well as for other small volume pipetting applications such as ELISAs. The unique geometry of the 50 μl tip makes it compatible with many different types of labware, including microplates, deep-well plates and sample tubes; it is also able to pierce septa. Its universal nose cone is designed for our existing Freedom EVO® series of liquid handling instruments and, in particular, for the recently launched 8 Plus 1 Access™ liquid handling arm for biopharmaceutical research and the HID EVOlution™ platform for forensic applications. Tecan Journal 2/2008
Our disposable tips are produced to meet critical technical specifications, maintaining high standards of quality and reproducibility. Each tip is individually inspected and tested during manufacturing. The validated production process meets the requirements of ISO 9001 and ISO 13485. All Tecan conductive disposable tips • minimize risk of carryover • reduce the chance of biological samples being contaminated (eg. RNase) • increase throughput by minimizing tip washing steps • enhance pipetting performance and results through optimized design • are available with or without filter. For more information on all our consumables products, visit our new consumables pages at www.tecan.com/consumables
For more information on the PowerScanner, visit www.tecan.com/PowerScanner, where you can also request updates and enter a competition to win a NanoQuant Plate™ (for Infinite® 200), LaserCheck™ (for LS Reloaded™), hybridization chamber set (for HS 400™ Pro) or an Apple iPod touch 16 GB. Apple and iPod are trademarks of Apple Inc., registered in the US and other countries.
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
PRO DUCT N EWS
www.tecan.com/get-journal
x 1000 wells per hour
Added functionality with the MultiChannel Arm™ 384 gripper option 0 100 0 9 7 1 0 3
A gripper for the recently launched MultiChannel Arm™ 384 brings greater functionality to the high throughput pipetting head, and will be available as an option or upgrade. The cost-effective alternative to a robotic arm mounts onto the MultiChannel Arm 384, and shares its structure and x axis. The gripper enables flexible access to all areas of the Freedom EVO® workstation with its unlimited 360-degree rotational axis, and the full range of its z axis allows the gripper to reach positions from the level of the worktable surface, up to
the highest shelf of a hotel. It is able to access objects located at the back of the instrument, such as labware stored in hotels and incubators, and can load/ unload plates from readers or hotels on the right hand side of the platform. The gripper can maneuver a plate by grasping either its short or long side. The gripper also minimizes the risk of sample contamination by removing the plate lid only for the duration of a pipetting step, replacing the lid immediately afterwards.
8 7
SUCCESSFUL RUNS
Scientific instrumentation. Not for use in human clinical or diagnostic procedures.
The Freedom EVOlyzer® takes routine clinical diagnostics tasks off your hands Tecan has launched an enhanced version of the Freedom EVOlyzer® for fully guided, automated processing of ELISAs through its new generation Freedom EVOlution™ 2.0 software, which offers an exceptionally user-friendly graphical user interface with optional touchscreen and a number of new application-oriented features. The open-platform, IVD-D compliant Freedom EVOlyzer automates ELISA kits from a variety of manufacturers and manages the entire workflow, from organizing the initial worktable configuration to providing the final assay results.
simple graphical loading dialogs and loading interface provide easy operation throughout the process, and its flexible scheduling options allow the order of assay processing to be defined and prioritized. Assays can also be bunched into profiles that facilitate everyday
routine tasks and the system is suitable for working in batch mode using worklists or via an ASTM connection. For more information on Tecan’s Freedom EVOlyzer, visit www.tecan.com/evolyzer
The Freedom EVOlyzer
The Freedom EVOlution 2.0 software performs many routine laboratory tasks, such as continuous sample loading and automated processing of pre-dilution factors. A volume pre-checking function ensures there are sufficient reagents for all scheduled tasks, allowing long walkaway times for the user with complete peace of mind. The software’s Tecan Journal 2/2008
5
and don’t forget
P ROD U CT N EWS
Sub microliter pipetting with Te-PS™ tips The Freedom EVOŽ workstation configured with Te-PS™ tips is an ideal platform for life science applications such as protein crystallography, where accurate pipetting in the sub microliter range is essential. The laser-guided Te-PS positioning system and specially-designed Te-PS tips have been developed for robust and accurate use of high density labware formats, such as 1,536-well microplates or high density protein crystallography plates.
#6Ă&#x;; =
Tests showed that the Freedom EVO workstation with Te-PS tips is capable of contact pipetting in the sub microliter range with very good precision and accuracy across the volume range of 100 nl to 1.0 Îźl, even for highly viscous liquids. The Te-PS tips achieved better precision and accuracy compared to the specifications of a standard manual pipette in this volume range, and achieved excellent consistency through each of the eight Te-PS tips. Dry contact pipetting performance was also shown to be of similar accuracy and precision to wet contact pipetting.
6OLUMEĂ&#x;;”L=
Research application, not for clinical diagnostic use. For more information, visit www.tecan.com/proteincrystallography
#6Ă&#x;; =
#6Ă&#x;; =
6
6OLUMEĂ&#x;;”L=
Fig 1 Precision of pipetting water with Freedom EVO workstation (green bars) and a standard manual pipette (blue bars). Data for the Freedom EVO workstation was obtained from gravimetric experiments using the Freedom EVO workstation with Te-PS tips. Data represent 480 data points derived from 10 measurements with eight tips in each of six separate experiments. Data for manual pipetting are specified performance parameters quoted for an adjustable manual pipette. Tecan Journal 2/2008
6OLUMEĂ&#x;;NL=
Fig 2 Precision of pipetting 40% PEG 10,000 using the Freedom EVO workstation and Te-PS tips. The data represent 480 datapoints derived from 10 measurements with eight tips in each of six separate experiments.
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
PRO DUCT N EWS
www.tecan.com/get-journal
Freedom EVOware®
Simple control of complex sample pipetting procedures The latest Freedom EVOware® Sample Oriented software, version 1.2, is now available for the Freedom EVO® family of workstations. This software enhances Freedom EVOware Standard and Freedom EVOware Plus with a number of samplebased pipetting, diluting and pooling functions. Freedom EVOware Sample Oriented is ideal for laboratories that need to perform complex pipetting and dilution sequences, and for those with variable joblists, assays and plate definitions. Freedom EVOware Sample Oriented is essential for pooling samples* with the Freedom EVO Clinical and TePoolSafe™, and controls primary pooling, sub-pooling, cross pooling and pipetting to archive plates. The plug-in wizards provide simple, graphically-guided operation for many tasks, including setting up assay plates, and defining pipetting sequences and pre- or serial dilutions. The wizards also manage sample replicates, control reagent addition and volume tracking,
and allow parallel pipetting of samples to one or more assays. The optional loading interface detects whether samples have been manipulated prior to pipetting and assists the user with loading of consumables onto the workstation. Positive identification of samples, plates and liquids can be carried out using PosID™ 2 or PosID 3 on the Freedom EVO 100, 150 and 200, or using a hand-held scanner on the Freedom EVO 75; sample identities can be tracked throughout pipetting steps and output files created. Freedom EVOware Sample Tracking is available as an optional add-on that creates printed reports for each plate (Sample Tracking Embedded) and provides long-term data storage (Sample Tracking Standard). Freedom EVOware Sample Oriented can be used with Magellan™, Tecan’s sophisticated data processing software for detection instruments, and has LIMS and ASTM E1381/E1392 connectivity.
The software package is included with the Freedom EVO 75, and can be ordered in addition to Freedom EVOware Standard or Plus for other Freedom EVO platforms. Freedom EVOware Sample Oriented allows a 30-day evaluation upon installation of Freedom EVOware on an instrument, before a dongle is required for continued operation. * Freedom EVO Clinical, pooling and Te-PoolSafe option not available in the USA For more information on Tecan’s Freedom EVOware Sample Oriented, visit www.tecan.com/freedomevowaresampleoriented
Making advances in regenerative medicine Tecan is collaborating with tissue engineering researchers at the University of Zurich to develop a novel, automated system for propagation of quality controlled cells that will be suitable for use in regenerative medicine therapies, such as intervertebral disk implants. The project is funded by the CTI, the Swiss Confederation’s innovation promotion agency, which supports the transfer of knowledge and technology between universities and businesses. The new solution will control all necessary cell culture steps, including the isolation,
seeding, proliferation, harvesting and analysis of cell lines and primary cells. Quality control will be maintained through two integrated detection devices that will determine the degree of cell confluence and detect the expression of cell type-specific genetic markers. The platform will be easily adjusted for culture of multiple cell types, providing standardized and affordable treatment procedures for regenerative medicine. Research application, not for clinical diagnostic use.
SEM (scanning electron microscopy) image of intervertebral disk cells seeded in a 3D collagen gel (gel surface) Picture supplied by Karin Wuertz, University of Zurich, Switzerland Tecan Journal 2/2008
7
and don’t forget
CO R POR ATE N EWS
Te-Care™ service contracts for peace of mind
The flexible range of Te-Care service contracts, which can be tailored to suit customers’ preferences and requirements, include the Te-Care Complete Contract, Te-Care Maintenance Contract and Te-Care Repair Contract.
M P L E TE
Te-Care
TM
Service Contracts
TE
The Te-Care Complete Contract optimizes the performance and uptime of customers’ systems with regular support, covers maintenance and repair and eliminates unexpected service expenses. The Maintenance Contract offers regularly scheduled product maintenance to reduce downtime and administration costs, while the Repair Contract extends instruments’ initial warranty by up to two years. Both the Complete and Repair Contracts offer quick response times, with priority treatment within 48 hours.
PA I R
CO MAIN
Tecan has launched Te-Care™ service contracts, offering a consistent service across the world to provide optimal reliability, lifetime and productivity of Tecan systems for all customers. The new global service contract structure gives access to a global network of experienced, certified field services engineers and applications specialists.
RE
8
NANCE
•
New look web content Tecan constantly updates its website www.tecan.com so that we can provide the most relevant, interesting and up-to-date information to you. Two major changes were made to the website in March as part of this continuing process, with launches of the new ‘consumables’ and updated ‘about Tecan’ sections. The new consumables pages www.tecan.com/consumables now list all of our available consumables, with detailed information on each individual product. The consumables are organized
Tecan Journal 2/2008
into four product groups: disposable tips, sample storage tubes (REMP), sample and reagent containers, and Cavro® accessories and consumables. The ‘about Tecan’ section www.tecan.com/about now contains many updated pages, including a company fact sheet, news pages, a fascinating history of Tecan, details of our quality policy and exciting job opportunities.
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
C LI N ICAL DIAGN O STI CS
www.tecan.com/get-journal
Tamine Hosseini (third from left) and colleagues in the Department of Clinical Chemistry at the Sahlgrenska University Hospital
Automated pre-analytical blood sample sorting The Sahlgrenska University Hospital in Gothenburg – one of the largest hospitals in Sweden – is saving considerable time using a Genesis FE500™ workstation for the pre-analytical processing of blood samples. At Sahlgrenska, the sample reception section of the Department of Clinical Chemistry is responsible for sorting and processing all hospital samples that arrive for laboratory analyses, including microbiology, immunology and serology, as well as the clinical chemistry samples. The laboratory’s Genesis FE500 workstation processes EDTA and citrate coagulation tubes for the full range of departments, as well as preparing some samples for archiving, automating the centrifugation and decapping of the sample tubes in preparation for subsequent analysis. “Before we had the Tecan system, we had to process these samples manually,” explained Tamine Hosseini, head of sample reception. “I would estimate that we are now saving around 30 to 40 per cent of the time originally spent in sorting, centrifugation and decapping.
The sample barcodes are scanned and registered into our laboratory information system (LIS) along with all the patient information and analysis requests. Once loaded onto the workstation, the barcode specifies the tests needed for each tube and the samples are sorted and processed accordingly.” “Our staff enjoys working with the Genesis FE500 workstation, largely because it needs virtually no daily, and only minimal weekly, maintenance work, compared to the analytical instruments, which require frequent calibration and extensive maintenance routines,” Tamine added. “We have had the Genesis FE500 system for one year and plans are now underway to use it for processing urine samples in the near future to make the most of the capabilities this instrument offers.”
For more information on Tecan’s Genesis FE500, visit www.tecan.com/genesisfe500
Tecan Journal 2/2008
9
10
and don’t forget
M I C ROP L ATE R EAD ER
Cost-effective PAMPA with Infinite® M200 Tecan and pION INC have combined pION’s PAMPA Explorer™ with Tecan’s microplate readers for drug permeability screening using the Double-Sink™ parallel artificial membrane permeability assay (PAMPA). pION, a biotechnology company located in Woburn near Boston, USA, specializes in developing laboratory products for screening and analysis of pharmaceutical compounds, in particular for determining solubility, dissolution, permeability and human absorption.
The pION team: (left to right) Oksana Tsinman, research chemist, Dima Voloby, scientific programmer, Dr Konstantin Tsinman, HTS group leader, Dr Alex Avdeef, CEO/CSO
“
The PAMPA software has been adapted to be able to take the raw data from the Infinite M200 and process it to generate the assay results.
”
Tecan Journal 2/2008
pION’s Double-Sink™ permeability assay measures a compound’s concentration on either side of the assay membrane, detected with a plate reader, in order to quantify the compound’s permeability, which is calculated by the PAMPA system’s software. The Double-Sink™ membrane is designed specifically to mimic the absorption characteristics of the human intestinal wall, and has been shown to correlate with actual human jejunal data. pION has since developed a number of products based on the assay, including a blood-brain barrier PAMPA system. “The PAMPA Explorer™ is an entry-level system that allows customers to use the validated PAMPA software and assay protocols with a microplate reader for fast and robust analysis in a manual setting,” said Dr Cynthia Berger, CFO at pION. The PAMPA Explorer™ now incorporates Tecan’s Infinite® M200 microplate reader for detection and quantitation of hundreds of compounds per day. The Infinite’s advanced quad4 monochromators™ technology provides excellent sensitivity and allows the user to select any wavelength from UV to NIR, and to perform absorbance, excitation and emission scans.
“The PAMPA software has been adapted to be able to take the raw data from the Infinite M200 and process it to generate the assay results,” explained Cynthia. “The Infinite plate reader was an obvious choice for the PAMPA Explorer™, because our market research discovered that this is the most popular detection system among our target customers. It was relatively straightforward to integrate the Infinite into the PAMPA, the development process went smoothly and we have very good communications with Tecan.” The PAMPA Explorer™ can be upgraded to a fully automated Tecan Freedom EVO® platform system when increases in sample throughput are required. Double-Sink, PAMPA Explorer and pION are trademarks of pION INC. Research application, not for clinical diagnostic use. For more information, visit www.tecan.com/adme
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
CO R PO R ATE N EWS
www.tecan.com/get-journal
Hotel Nikko Tokyo in Odaiba, overlooking Tokyo Bay. The name ‘Odaiba’ originated from the late Edo period (1603-1867) and derives from the government-built artillery platforms (daiba) to protect Edo, the old Japanese capital that is now Tokyo, from attack by foreign ships. Photo provided courtesy of Hotel Nikko Tokyo
Asia Pacific Sales Meeting in Japan Tecan’s Asia Pacific 2008 Sales Meeting was held from February 18 to 19, at the Hotel Nikko (Japan Airlines) Tokyo in Odaiba, located in the Tokyo Bay area. The annual sales meeting, co-hosted by Tecan Japan and Tecan Asia Pacific, brought together Tecan’s regional offices and sales agencies, and was attended by participants from Australia, China, India, Korea, Malaysia, Singapore, Taiwan, Thailand and Japan. The meeting was opened by Carl Severinghaus, Senior Vice President, followed by a review of 2007 and future plans for 2008 in Asia Pacific and Japan and summaries by managers of each business unit. There were announcements and information about new products from head office representatives and, with a presentation from each regional office and sales agency, it was an excellent opportunity to exchange information among the Asian countries. Five section meetings were held for liquid handling, detection, REMP sample management, components, and service and consumables, during which the opportunity for handson training for new products was enthusiastically received. On the evening of the first day, attendees enjoyed an evening boat cruise and Japanese cuisine such as tempura,
sashimi and chanko-nabe (one-pot stew), gazing upon the night view of the seaside city sub-center from a yakatabune – traditional Japanese cruising boat – floating in Tokyo Bay. Further entertainment during the feast came from performances by dancers, called ‘furisode-san’ (‘furisode’ are long sleeved kimono, traditional Japanese formal attire), and the evening was perfect for members of the team to catch up with each other.
A section meeting in progress
Attendees at the Asia Pacific 2008 Sales Meeting
‘Furisode-san’ entertained on the first evening Tecan Journal 2/2008
11
12
li quid handling AN D ROB OTICS
and don’t forget
Automated differentiation of stem cells with the Freedom EVO® and Millicell®-24 filter plates Scientists at Millipore have developed an automated method for the neural differentiation of embryonic stem cells (ESCs) in monolayers, instead of the non-automatable, three-dimensional cell clusters that are typically used. The approach relies on a Freedom EVO® 150 liquid handling platform from Tecan, with Millicell®-24 co-culturing filter plates from Millipore.
Millipore Corporation is a global life science company, headquartered in Massachusetts, USA, that provides technologies, tools and services for bioscience research and biopharmaceutical manufacturing. Millipore has worked with Tecan for a number of years in a variety of laboratory automation projects and many of Millipore’s consumable devices are automation-friendly. In this latest collaboration, Libby Kellard, automation specialist at Millipore, and her colleagues have used a Freedom EVO 150 workstation to develop an automated method for the expansion and neural differentiation of mouse embryonic stem cells (mESCs). ESCs are widely used in a growing number of laboratory research areas, and successful differentiation of cultured ESCs into specific tissues is key to applications in tissue engineering, drug discovery and regenerative medicine. ESCs are usually co-cultured with mouse embryonic fibroblasts during expansion to maintain the ESCs in an undifferentiated state; removal of the fibroblasts is necessary to initiate differentiation. Although culturing of ESCs has been successfully automated in the past, manipulating the differentiation of these cells is not conducive to automation and is typically done manually. Differentiation protocols are labor-intensive and commonly include the use of three-dimensional cell clusters (embryoid bodies) that allow the initial formation of the three germ layers, which develop later into different tissue types. An alternative method has been proposed where ESCs grown as a monolayer can differentiate into neurites under the right Tecan Journal 2/2008
Fixed tips on the liquid handling arm add media to cultured cells in the Millicell®-24 filter plate
conditions.1 The scientists at Millipore adopted this approach to develop an automated technique for stem cell differentiation using the Freedom EVO workstation with Millipore’s Millicell®-24 filter plate. Monolayers of ESCs are indirectly co-cultured with a feeder layer of mouse embryonic fibroblasts, separated by a 1 μM PET membrane that allows easy removal of the fibroblasts to initiate differentiation.
biology devices at Millipore. “Cells can be grown on the membrane, allowing researchers to access both the top and bottom (apical and basolateral) surfaces of the cells. This cannot be done with standard solid plastic cell culturing plates. The microporous membranes are available as single inserts or in 24- and 96-well single units, suitable for high throughput screening applications, including automated workflows.”
“We developed the original Millicell® product in the early 1980s as a novel cell culturing device with a microporous membrane support,” explained Ken Ludwig, group product manager for cell
“The first step was to establish the cell culturing method and expansion using the Freedom EVO,” Libby Kellard said. The Freedom EVO includes a specially integrated Liconic incubator with an
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
li q uid handling AN D RO B OTI CS
www.tecan.com/get-journal
Neurite Differentiation Automation
Manual
Neurofilament
Nestin
The Freedom EVO workstation is housed within a HEPA filter hood to minimize contamination
automatic shuttle that brings plates to the deck for the robotic manipulator (RoMa) arm to pick up. Other modules include a tilting rack on the deck that aids removal of media from the feeder wells by the 8-channel liquid handling arm, and cooling and warming racks for the media. The entire platform is housed within a HEPA (high efficiency particulate air) filter hood to minimize contamination. “The biggest challenge was getting all the equipment fitted, because the engineers had to build the system within the HEPA hood,” Libby continued. “It was important to ensure that the coculturing environment meets cleanroom requirements before we could grow stem cells. We validated the system using standard cell lines such as Caco and MDCK. The people from Tecan were really helpful; they set up the entire system for me and helped with all the initial programming so that if any modifications needed to be made later, I was able to do them myself.” “We found the people at Tecan really easy to work with,” Ken added. “They seem very much focused around the customer and the application, and that really helped.” “We have now shown that mESCs can be successfully maintained by the Freedom EVO in an undifferentiated state using the Millicell®-24 filter plates for indirect
Beta Tubulin 2G10
mESC cells were expanded for 4 days and then allowed to differentiate for 15 days using the Tecan Freedom EVO workstation. Plates that were handled by automation behaved similarly to plates processed manually. All plates stained positive for Neurofilament, Nestin and Beta Tubulin 2G10. In both cases Neurite differentiation occurred starting with a cell monolayer as opposed to embryoid bodies which is the typical way of producing neurites.
co-culturing, where mouse embryonic fibroblasts are seeded in a layer that is separated from the mESCs by the 1 μm PET membrane. The filter membrane makes it easy for the Freedom EVO to automatically remove the fibroblast layer, causing cell differentiation, and we achieved successful differentiation of the cells along neural lineages,” said Libby. “There were no discernible differences in cells that were automatically cultured and differentiated when compared with cells that were manually prepared at the same time. We have not yet optimized the program for high throughput but, at the moment, it certainly avoids the need for anyone to come in and feed the cells at the weekend! The system takes care of all the routine culturing steps without any intervention.”
The method is likely to be suitable for all mouse ESCs and could potentially be adapted for other ESC types. “I think it would benefit any stem cell research application where scientists are trying to maintain ESCs over a lengthy period of time,” Libby concluded. Reference 1. Pachernik J, Esner M, Bryja V, Dvorak P, Hampl A (2002). Neural differentiation of mouse embryonic stem cells grown in monolayer. Reprod Nutr Dev 42: 317-326 Millicell and Millipore are trademarks of Millipore Corporation or an affiliated company. For more information on Tecan’s Freedom EVO workstations, visit www.tecan.com/freedomevo
Tecan Journal 2/2008
13
14
and don’t forget
WAS H E RS
Developing one-step allergy testing panels using the HydroFlex™ system VBC-Genomics in Vienna uses a Tecan HydroFlex™ system for automated washing of its microarrays for allergy testing. The arrays are spotted onto VBC’s proprietary surface, segmented into a 96 well format with a Whatman FAST® frame slide adapter and then processed by the HydroFlex, which halves the slide processing time and improves the signal quality and consistency of results.
The team at VBC-Genomics (left to right): Rene Ertler, Tamara Braun, Sabine Hutter, Claudia Kirisits, Marianne Brill and Christian Harwanegg
The Tecan HydroFlex system
Tecan Journal 2/2008
VBC-Genomics Bioscience Research was founded in Vienna, Austria, in 1999, as a spin-off company from Vienna University, and was established for providing microarray-based genetic research and screening tools. The company currently focuses on developing microarrays to screen for large numbers of allergens, in order to identify the specific triggers of individual patients’ allergic reactions. VBC-Genomics has already launched a panel of 89 different allergens that allows rapid, one-step allergy testing from a single drop of blood; the latest version of this product, which will be called ImmunoCAP® ISAC® (Immuno Solid-phase Allergen Chip), will be launched later this year in co-operation with Phadia, Sweden. Phadia is the global market leader for in vitro allergy diagnosis and one of VBCGenomics’ major shareholders.
“We have been keen to automate our array development procedures for some time, and we chose Tecan because it is important that the processes meet certain quality standards,” said Christian Harwanegg, COO at VBC-Genomics. “The arrays are CE-marked, registered medical devices that meet IVD-D requirements, and they are spotted with highly standardized, purified or recombinant allergen molecules, rather than the extracts used by many other companies.” These arrays can be used by healthcare professionals to test patient samples against a broad range of molecules that may be responsible for their allergies. The information can be used to define more suitable, individualized treatment or avoidance regimes. ISAC® allows semiquantitative analysis of IgE
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
WASH ERS
www.tecan.com/get-journal
application specialist was able to create a customized plate setting for a 64-field format for the washer so that we can wash four slides simultaneously using our Whatman slide adapter.”
ImmunoCAP® ISAC® enables rapid, one-step allergy testing from a single drop of blood
immunoglobulins from human serum and is the first biochip-based allergy test approved for allergy diagnosis according to EU regulations. The arrays contain four different segmented chambers, each of which can contain up to 400 individual allergens tested against just 20 μl of human serum. Any IgE antibodies – indicating an allergic reaction – bind specifically to their respective allergens spotted on the array, emitting fluorescent signals that are automatically identified using standard microarray scanning techniques. “Our panels aim to cover as many relevant allergens as possible in a single assay, because even if you know that 90 % of rhinitis patients, for example, react only to specific antigens, it is important to cater for the additional 10 % so doctors can be confident that they have identified all of a patient’s allergy triggers,” Christian explained. “We chose the HydroFlex because we needed a system that is affordable, practical and easy to integrate into other laboratory automation platforms and workflows. We looked at several instruments initially but we were convinced that the HydroFlex was the only system that was able to meet all of our needs, including its compliance with the IVD directive 98/79/EC in ELISA configuration, flexibility and software, without any compromise on our part. Its flexibility is essential to us, as Tecan’s
“We now rely on the HydroFlex for many of our internal regulatory testing and product development procedures, the automated washing, shaking and mixing steps are very important in eliminating human error so that we can truly standardize the testing processes. We are really impressed with the performance of the HydroFlex; it has already helped to speed up our assay processing time from around five hours to just two hours including all washing steps. We have significantly increased our throughput so
that one person can easily process about 200 tests (representing almost 20,000 results) in a day, requiring minimal handson time. Perhaps the most important benefits we have seen are the increases in both the sensitivity of the assay and in its reproducibility, each by at least 50 %. In the future we hope to integrate the HydroFlex system with an automated liquid handling platform, such as the Freedom EVO®, to fully automate our laboratory processes.” FAST is a trademark of companies within the Whatman Group. ISAC is a trademark of VBC-Genomics. ImmunoCAP is a trademark of Phadia AB. For more information on Tecan’s HydroFlex, visit www.tecan.com/hydroflex
Preparation of allergen molecules
Tecan Journal 2/2008
15
16
and don’t forget
C L I N I CA L D I AG NOSTICS
Efficiency by automation – high throughput sample processing The diagnostics laboratory at the Catholic Medical Center (CMC) in Manchester, New Hampshire, USA, has chosen the Genesis FE500™ platform to automate sample handling, successfully turning its ‘stat lab’ into a full service laboratory and processing three times the previous volume in the same laboratory space.
(left to right) Pat Felton, Steve Harkins, Roberta Provencal and Krysti Pinard with the Tecan platform
CMC’s diagnostics laboratory previously handled only in-patient blood samples and outsourced much of its out-patient blood testing. The hospital is building a new, purpose-built full-service blood diagnostics laboratory due for completion by the Spring of 2009 and, in anticipation, the laboratory has started processing the huge number of outpatient samples in-house. Roberta Provencal, Director of Laboratory Services, explained: “We recently converted from a stat lab to a full-service laboratory, covering a number of services including blood banking, chemistry with immunoassays, hematology, serology, mycology and microbiology, by completely restructuring the way the samples are handled. Now we also receive all the outpatient samples from the hospital’s lab collection station as well as various collection sites around the community, which has tripled sample throughput. We couldn’t really do anything to increase working area until the new facilities are complete, so we had to make the most of every inch of available space within our existing premises.” “I had previously worked with a Tecan system for several years where it saved us
Tecan Journal 2/2008
so much personnel time, and I knew that it would be the perfect solution. Without Tecan’s Genesis FE500 workstation, we would have to use a bank of centrifuges, and employ many people to manually process the specimens – we simply didn’t have the room to work that way.” “With the current system, samples are assigned a barcode and entered into the laboratory information system (LIS) during collection, which has eliminated the potential for manual errors in sample handling,” continued Roberta. “The barcoded sample tubes arrive at the specimen processing area in our laboratory, and all we need to do is put tubes onto the Genesis FE500 system. The barcodes contain information about the tests that have been ordered for the samples, and the Genesis FE500 can process each sample, placing each tube into frozen, refrigerated or ambient output lanes as appropriate. Samples can be processed immediately with this set-up, and this has both decreased the risk of handling errors and improved our specimen integrity. We can now have routine test results within three hours of receiving the specimens.”
(left to right) Kristi Pinard, Muriel Chabot, Doug Cloutier and Pat Felton, with the Genesis FE500
“The system is easy to use and maintain. The decapping feature is popular with our technicians, because it has minimized manual handling of specimens and aerosol exposure of personnel, and removed the tedious need to manually uncap large numbers of tubes.” Roberta concluded: “Tecan set up the Genesis FE500 for us and the workstation was operational in just one week – the Genesis FE500 system is the best thing since sliced bread!” For more information on Tecan’s Genesis FE500, visit www.tecan.com/genesisfe500
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
C LI N ICAL DIAGN O STI CS
www.tecan.com/get-journal
High throughput EIAs with Freedom EVO® Clinical Laboratory services company synlab offers analysis of human, animal and environmental samples and, at its Augsburg site in Germany, it has chosen two Freedom EVO® Clinical workstations in combination with the BEP® III system from Siemens to process an enormous throughput of EIAs to diagnose infectious diseases.
Close-up of one of the Freedom EVO 150 Clinical workstations
Clinical analyses are performed in 55 synlab locations in Germany and 18 abroad by competent teams of doctors, scientists, engineers and skilled laboratory personnel, who ensure that the analyses are carried out to the highest standards. At the Augsburg site in Germany, the serology department performs routine immunology assays of serum and liquor samples. The laboratory runs more than 124,000 EIAs a year and uses two Freedom EVO 150 Clinical workstations to pre-process the assays for analysis on the Siemens BEP® III system, an automated microplate processor.
(left to right) Petra Müller, Manuela Strack and Dr Ulrike Hauser, of the Augsburg serology department
Using Novagnost® and Enzygnost® tests from Siemens, a wide variety of assays can be run simultaneously on the Freedom EVO Clinical, using a different strip on the microplate for each assay. Nearly 50 different parameters have been implemented for various infectious agents, including Epstein-Barr virus, Herpes simplex virus, measles, adenovirus, influenza, Helicobacter pylori, periodontitis, Chlamydia, Candida and Mycoplasma. “In collaboration with Siemens, specific software features were implemented in Freedom EVOlution™ 1.9 to accommodate the Siemens tests and allow data to be transferred smoothly to the BEP® III system,” elaborated Caroline Huber, Tecan’s product manager for the Freedom EVO Clinical. “Liquor samples require special pre-dilution steps, for which the Freedom EVOlution 1.9 has also been programmed to process appropriately.” The Freedom EVO Clinical workstations have an integrated liquid handling (LiHa) arm with four channels using standard tips, and are also equipped with a wash station and PosID™3. The automated platforms process samples from 3 ml and 10 ml tubes to perform the assays in 96-well strip plates.
The Augsburg serology department is led by Dr Ulrike Hauser and senior technicians Petra Müller and Manuela Strack. “We have already been using automated systems for 10 years in our laboratory, but the old system was getting outdated and its capacity was becoming insufficient for our needs,” explained Manuela. “We chose the Freedom EVO because of its flexibility, which gives it the ability to adapt to our many different test formats. The Freedom EVOlution 1.9 software is easy to use and has a range of valuable features, allowing us to choose from a variety of assays for each patient sample. The graphic user interface makes handling very simple, and the workflow can be clearly arranged and easily followed. Fully automated sample processing and high throughputs were essential requirements so, after the set-up issues were resolved, the Freedom EVO Clinical workstations have been particularly beneficial to the laboratory.” BEP, Enzygnost and Novagnost are trademarks of Siemens Healthcare Diagnostics. For more information on Tecan’s Freedom EVO clinical, visit www.tecan.com/freedomevoclinical Tecan Journal 2/2008
17
18
and don’t forget
A P P L I CATI ON B IOP HARM A
Automated pH measurements and solubility screening of candidate compounds in pharmaceutical development Scientists at AstraZeneca UK in partnership with Tecan are developing a fully automated solubility screening platform, a Freedom EVO® 200 liquid handling workstation integrated with a tailor-made data capture system, a novel pH flow cell carrier and a small footprint HPLC system.
AstraZeneca’s Early Development Group, part of Pharmaceutical Research and Development at the Alderley Park site in Cheshire, UK, receives many compounds made or isolated by AstraZeneca’s Discovery Group and assesses them for their suitability for development. This requires large-scale solubility screening at variable pHs and, until recently, this was performed using time-consuming and costly manual processes. AstraZeneca recognized that, to improve efficiency, automated solubility screening was needed and ideally should require no operator intervention once the screen has begun. Suitable fully automated systems were not available off-the-shelf at that time and it was then that the scientists approached Tecan. Rod Kittlety, Senior Research Scientist in the Early Development Group, explained, “We had a number of criteria for our automated solubility screening system. It needed to measure solubility at different times and over a large dynamic range (1 µg/ml to more than 20 mg/ml) and the pH to an accuracy of 0.1. A system using centrifugation rather than filtration for solid separation was essential, because solubility measurements at concentrations below around 50 µg/ml can be affected by compound-specific absorption to filters. Another requirement was the incorporation of an HPLC system which could handle a number of different methods thus allowing the stability of the compound in question to be assessed during the experiment. We wanted to Tecan Journal 2/2008
eliminate operator intervention after the process started, and to be able to control the temperature over a range from 4 ºC, which is the minimum storage temperature, to 37 ºC, the physiological temperature. Tecan worked with us to develop a system that satisfies all these requirements.” AstraZeneca’s solubility screening platform consists of a Freedom EVO 200 workstation equipped with a specially-designed automated pH measuring device and integrated with liquid handling (LiHa) and robotic manipulator (RoMa) arms, an automated centrifuge, and a small-footprint HPLC system. The flow-through pH cell is controlled by software that integrates with Freedom EVOware®, allowing calibration, measurement and pH stability analyses. An integral data capture system converts the PC into a configurable data acquisition device for automated calibration and measurement of pH, then returns the captured data to Freedom EVOware for export. The benefits of this adaptation include stable pH readings and correction of sample pH readings with calibration data, in 96- and 384-well formats. For most compounds, the amount of substance needed for analysis is small; two 50 µl aliquots of diluted sample injected sequentially give good data for most buffers. “The equilibrium solubility of a compound increases as it starts to ionize, and this is dependent on pH, so
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
APPLICATIO N B IO PHA R M A
www.tecan.com/get-journal
measuring the pH of the suspension accurately is essential when assessing the pH-solubility relationship,” said Rod. “We needed this adaptation to obtain all these pH measurements at defined time intervals. We use four buffers with defined pHs from 1.68 to 10.0 to calibrate the electrode which means we can cover the physiological pH range within which pharmaceutical formulations are required to work. It is important to measure substance solubility over a sufficiently wide range of pHs so that solubility under physiological and formulation conditions can be assessed.” The pH calibration is done before each solubility time point. The solubility screening platform allows sample pH measurements to be carried out while sample aliquots are centrifuging. The centrifuged samples are then diluted and aliquots of these are injected into the HPLC system; the solubility of 36 samples at three time points is measured usually up to 48 hours, and many pH measurements are automatically performed during the process. “Solubility results from test compounds from the automated system agree very well with those from manual screens,” Rod continued. “The automated system gives us the precision we need over the required pH and solubility ranges, and the instrument is very robust. I have done about 3,000 sample injections on the pH system while setting it up
Rod Kittlety loads the carrier which holds the pH calibration buffers
and it works really well. The system has a great deal of potential for allowing us to do many of the things we normally do in the laboratory, but automatically and in a smarter way. For example, we are now using the system for a plasma precipitation screening assay to measure the level of active compound in plasma samples, and we can complete this in just half an hour using the Freedom EVO platform, compared with almost a whole day when performed manually.” Research application, not for clinical diagnostic use. For more information, visit www.tecan.com/analytics
Tecan Journal 2/2008
19
20
li quid handling AN D ROB OTICS
and don’t forget
True high throughput compound management for Galderma French pharmaceutical company Galderma specialises in the development of drugs for a variety of dermatological complaints, including acne, rosacea and psoriasis, and has chosen a customized Freedom EVO® 200 liquid handling workstation to manage its huge library of compounds used in high throughput screening.
Dr Bernard Schoot
The Structural Research Group at Galderma, headed by Dr Bernard Schoot, carries out drug development research using various proteomics methods, like protein crystallography, as well as techniques such as NMR for protein ligand interaction studies. “Within our department we have a Compound Management Group which is responsible for the registration, storage and distribution of all compounds that are produced and analyzed in our research,” explained Bernard. “We store compounds as both powder and in solution in our global storage platform, and we needed a compact, fully integrated workstation to maintain a complete library of all the compounds produced, analyzed and purchased by Galderma. From the start we wanted to configure the workstation to be dedicated to certain operations. Catherine Comino and Hamid Ait-Oufkir from our Compound Management Group carefully studied and defined all our workflows so we could accurately determine the functions and specifications required of the workstation. This information was then passed on to Tecan and the system was customized accordingly for us by reprogramming the software. As a result, we need to vary only a limited number of parameters to be able to do anything we want to do.” “Our Tecan platform is involved in the very early stages of drug development; preparing, storing and distributing the samples that are used for high throughput screening,” Bernard continued. “We chose a Freedom EVO 200 workstation, integrated with one
Tecan Journal 2/2008
liquid handling (LiHa) arm, one robotic manipulator (RoMa) arm and a Z8 pipetting arm, a balance, barcode readers for plates and vials, and a capping and decapping station. Plates can be loaded into a hotel and labeled with a barcode, and individual tubes within the plates also have a code that can be read from the bottom. The barcodes give access to all the information about the compounds, as well as the location of the tubes and the amount of the sample. All data are kept in the inventory in our laboratory information management system.” “Importantly, Tecan was able to satisfy a special requirement for compound storage in our laboratory, whereby the capped tubes are flushed with dry nitrogen before anything is stored in them and then, once the tubes are closed, they are not opened again until required,” continued Bernard. “This is because the dimethyl sulfoxide (DMSO) we use as the solvent is hygroscopic, and some of the products we store are sensitive to oxygen. This was a significant reason for choosing Tecan, together with the fact that we had access to a working model for testing.” Hamid, the technician responsible for the storage platforms, added: “Tecan was the only company we spoke to that was able to perform the whole of our workflow on one machine, which was very important because it made more efficient use of our bench space in the lab.” Catherine also commented: “We visited the factory several times to see the working model and, even after we chose Tecan, we continued thorough
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
www.tecan.com/get-journal
li q uid handling AN D RO B OTI CS
(left to right) Hamid Ait-Oufkir and Catherine Comino of the Structural Research Group at Galderma
discussions about the workflow and the exact requirements we wanted from the instrument. At this time, technical support from Tecan’s local team in France was very helpful.” Bernard is impressed with the throughput and capabilities of the Freedom EVO workstation: “From plates with stock solutions, we can make a double copy of 1,600 samples within 24 hours. We can also weigh compounds. Each capped, barcoded vial is tared on the workstation and, once compounds are analyzed, they are delivered to us in those vials. The vials are weighed again and then automatically, depending on what has been requested, the workstation dissolves and distributes the compounds into storage vials and tubes as necessary, after which the workstation produces
a report. All this information is included in the informatics of the system and you can automatically download it into an inventory, from which researchers can request the products. Everything is completely integrated now, but before we had the automated platform it would have taken a long time to do anything; in fact, we couldn’t do what we do now.” Hamid concluded: “We are happy with the Freedom EVO platform; the system is good, and we’re very happy with its performance.” Scientific instrumentation. Not for use in human clinical or in vitro diagnostic procedures.
“
Tecan was the only company we spoke to that was able to perform the whole of our workflow on one machine.
”
For more information on Tecan’s Freedom EVO workstations, visit www.tecan.com/freedomevo
Tecan Journal 2/2008
21
22
li quid handling AN D ROB OTICS
and don’t forget
Automating protein production processes for drug discovery at Bayer Schering Pharma AG One of the many functions of Bayer Schering Pharma’s Berlin research facility is to provide recombinant protein tools for the company’s lead discovery process. Scientists there have integrated two Freedom EVO® workstations into the protein production process, ranging from cloning and expression to purification, and including automation of GE Healthcare’s MultiTrap® 96-well plates.
Selected hits are then co-crystallized with crystallization-grade protein batches to identify the binding mode and to allow structure-based optimization. Our research covers a variety of applications, but we have particular experience with human drug targets, such as kinases and nuclear steroid receptors. For instance, the kinase projects aim to replicate the success of Nexavar®, a b-Raf kinase inhibitor, which is approved for renal cancer and is a potential pan-tumor drug.”
Technician Mario Mann loads samples onto the Freedom EVO 200
Bayer Schering Pharma AG, part of Bayer AG, develops new drugs and innovative therapies for improving healthcare in a range of specialist fields, with a research focus on oncology, cardiology, women’s health and diagnostic imaging. Identifying and evaluating potential new drug candidates is a vital aspect of lead discovery, and the company’s research center in Berlin supplies the protein tools that are essential to these processes. Tecan Journal 2/2008
“We perform more than a thousand protein expressions every year in order to deliver highly active protein preparations for high throughput screening and, in particular, to provide proteins for crystallization,” said Dr Dieter Moosmayer from the Protein Supply function at the Bayer Schering Pharma research facility in Berlin. “The protein preparations are screened against our compound library to identify small molecule inhibitors.
Being responsible for the whole protein production process, ranging from cloning of variants and mutations through to their expression and preparative purification, requires stringent analysis and quality control at every stage of the process. In order to deliver proteins within standard timeframes, the team switched to a multi-parallel approach a few years ago that included a Tecan Genesis liquid handling workstation. Dr Beate MüllerTiemann explained, “To go from the initial cloning stages through to crystallization of the protein can take a couple of months. We do not have the time to fail, so we used a strategy of miniaturization, parallelization and automation that helped us to find the optimal approach for each individual target gene within given timeframes.” “A library of expression variants is initially developed by a team of structural
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
www.tecan.com/get-journal
biologists and biochemists. These variants are then expressed in small-scale multiparallel expression systems and the best candidates proceed to purification, where downstream processing of preparative multi-step chromatographic purification is initiated, again in a multiparallel fashion, and finally up-scaled for structural determination.” The laboratory has recently upgraded its automated platform to include two Freedom EVO liquid handling workstations equipped with Te-VacS™ vacuum separation modules. “We decided to use Tecan instruments for these applications for two reasons,” said Dr Arndt Schmitz. “Firstly, Tecan technology has an impressive history of compatibility with kits and procedures that are relevant to our applications and, secondly, the instrument software is very flexible, allowing integration with equipment from other manufacturers,” he said. “The technical team at Tecan Germany is very knowledgeable and always available. Whenever we plan an upgrade, new script, or routine maintenance, the team is just a telephone call away. In addition, the training courses offered greatly aided our technicians Mario Mann, Ina Künkel, Andrè Hilpmann and Norbert Otto in setting up the automation here.” The laboratory’s Freedom EVO 150 is dedicated to automating molecular biology processes, including complete
Fig 1A. To examine cross-contamination, clarified E. coli lysate containing histidine-tagged enhanced green fluorescent protein ((His)6-EGFP, Mr 28 000) or histidine-tagged kinase ((His)6kinase, Mr 43 800) were applied in a chessboard scheme across one His MultiTrap™ HP plate, using a Tecan liquid handling workstation and the Te-VacS vacuum separation module. K: (His)6-kinase E: (His)6-EGFP.
li q uid handling AN D RO B OTI CS processing of the Invitrogen Gateway® recombinant cloning technology, and preparation of DNA minipreps. “The workstation is able to support complex chains of scripts such as Gateway® cloning, transforming and plating. That openness and flexibility was of particular importance to us,” Dr Schmitz said. “Performing thousands of tests on genetically modified organisms every year is an enormous task in terms of producing clones, transfection, purification and analysis.” The laboratory’s Freedom EVO 200 has an integrated GENios™ microplate reader and automates protein purification steps, including processing MultiTrap™ plates from GE Healthcare, which are ready-to-use 96-well filter plates for high throughput screening, enrichment and purification of tagged proteins from cell extract before quantification. Fellow staff scientist Dr Guido Malawski initiated beta-testing of the MultiTrap™ plates on the Freedom EVO and found that automated processing of E. coli lysates containing HIS- or GST-tagged target proteins provided a highly reproducible yield of target proteins, with no significant cross-contamination. The high throughput expression and purification of kinases with MultiTrap™ plates gave increased protein production throughput for X-ray crystallography. Screening of expression and purification at an early stage minimizes the workload
Fig 1B. Coomassie®-stained SDS-PAGE analysis (reduced conditions) of purified (His)6-kinase and (His)6-EGFP using His MultiTrap™ HP and a chessboard sample application scheme (Fig 1A). No significant cross-contamination was found. Lanes: M: molecular weight markers K: (His)6-kinase (Mr 43 800) in lysate and eluates of 12 wells E: (His)6-EGFP (Mr 28 000) in lysate and eluates of 12 wells.
by identifying the most interesting expression variants on which scaled-up purification can focus1. Both Freedom EVO workstations have quickly become an important part of the facility’s workflow. “We regard these instruments as our ‘co-workers’ and have even given them names,” Dr Schmitz confessed. “The Freedom EVO 200 is named Marvin while our molecular biology robot is called Brad Pitt! We have implemented standard operating procedures and protocols with these systems, which allows everyone in the department to use them.” He concluded: “Even today, we have not yet fully realized the potential of the systems; there is still plenty of room for growth.” Reference 1. Malawski GA, Hillig RC, Monteclaro F, Eberspaecher U, Schmitz AAP, Crusius K, Huber M, Egner U, Donner P, MüllerTiemann B (2006). Identifying protein construct variants with increased crystallization propensity - a case study. Protein Sci 15: 2718-2728 MultiTrap is a trademark of GE Healthcare Biosciences. Nexavar is a trademark of Bayer AG. Gateway is a trademark of Invitrogen Corporation. Coomassie is a trademark of ICI plc. For more information on this application, visit www.tecan.com/proteinpurification
Figure 2. SDS-PAGE analysis of eluates of histidine-tagged truncated variants of kinase, expressed in a transient insect cell system for 72 h. Lanes 06-31: histidine-tagged variants of kinase, E: EGFP (untagged transfection positive control), C: non-transfected Hi5 cells, B1 to B4: 0.5, 0.75, 1.0 and 1.5 μg BSA, respectively.
Tecan Journal 2/2008
23
24
and don’t forget
LIQ U I D H A N D L I NG
Automated screening systems in plant biotechnology Advanced Technologies Cambridge (ATC), a plant biotechnology company located on the Cambridge Science Park, UK, has chosen the Tecan Freedom EVO® 100 liquid handling workstation to perform complex pipetting tasks for setting up PCR screening assays.
ATC scientists (left to right), Rob Hurst, Matt Hope and Tim Beddoes
ATC was spun out from Twyford Plant Laboratories in 1988 as a wholly-owned subsidiary of British American Tobacco (BAT), and has in past years focused heavily on carbohydrate research, primarily to improve processing characteristics of potatoes and other crop plants. ATC has diversified into plant biotechnology and in vitro toxicological testing, with about 40 scientists involved in these two areas of research. The researchers at ATC enjoy a varied work environment, with opportunities Tecan Journal 2/2008
within tissue culture, molecular biology, biochemistry, genomics and toxicology. The toxicological studies at ATC use cell-based models to study the effects of smoke condensates without using animals in their testing, while the plant biotechnology section is currently working on projects including harm reduction and improved agronomic traits in tobacco. “We are developing projects on harm reduction,” explained Tim Beddoes, molecular biology technical specialist in
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
SERVIC ES AN D CO N SUM A B LES
www.tecan.com/get-journal
“
Our previous automated system was seldom used because it was not user-friendly.
”
the plant biotechnology section. “There are a number of toxicants in cigarette smoke with the potential to cause disease. These fall into several classes, for example nitrogenous compounds. The aim of the projects is to find ways to alter pathways within the plant that produce those compounds or their precursors. As part of these projects, transgenic plants are produced using a plant binary transformation vector to introduce genes of interest and common sequences. That common sequence can then be used to detect the presence of the introduced gene and determine how many copies of the gene have been introduced into those plants – a kind of quality control tool. In this way we can produce plants with defined genetics, which is essential for our plant breeding program and biochemical studies.” “We use real-time quantitative PCR (QPCR) to assay for the introduced sequences, and our Tecan Freedom EVO 100 workstation sets up the reaction plates for the assay. We used to perform all of our pipetting work manually, but now run several thousand assays per month on average. Managing this throughput was one of the reasons we decided to automate the process and, at the same time, it was also important to free up people’s time to do other things as much as possible. We are using a 96-well format for this process at the moment, but plans to switch to a 384-well format in the near future have made automation essential. The Freedom EVO 100 is also being used in the optimization of QPCR assays
where complex pipetting is involved, and may in future be used for some biochemical assays.” “Our current set-up has an 8-channel liquid handling arm and uses the 50 μl disposable tips which have been developed by Tecan. We perform our QPCR in final reaction volumes of 25 μl, because using relatively small volumes obviously cuts down on reagent and sample costs, and the 50 μl tips are perfect for pipetting the 5 μl of DNA and 20 μl of master mix. The 50 μl tip is also ideal for aliquoting fractions for experimental replicates.” “We have now optimized our method for setting up the reaction plates on the Freedom EVO with these tips. Test assays have confirmed pipetting accuracy, which we’ve been very happy with. The reproducibility is comparable to manual pipetting, but the tasks are performed much more quickly.” “Our previous automated system was seldom used because it was not userfriendly. I visited other laboratories to see how they used automated systems, and received a lot of personal recommendations for the Freedom EVO, including from colleagues at ATC who had used Tecan workstations at their previous place of work.” “We have had the Freedom EVO workstation for less than a year so far, and we are still developing and optimizing the system, so haven’t really pushed it to its full ability yet. We realize that there are still plenty of things that the system is capable of, and we are happy in the knowledge that we have that capacity for modification to handle new requirements in the future,” Tim concluded. For more information on all our consumables products, visit our new consumables pages at www.tecan.com/consumables Tecan Journal 2/2008
25
26
and don’t forget
QUA L IT Y
Ahead of the game for quality processes A recent article in the January 2008 issue of Qualität und Zuverlässigkeit (Quality and Reliability), the most widely read quality journal in German speaking countries, described a new quality management approach with a radically different organizational structure. A similar structure, simplifying QM and making it more efficient, has already been implemented at Tecan Austria in 2006, clearly showing that we are leading the way in the area of quality management.
At Tecan Austria, quality functions are integrated as a part of the whole process and not enclosed within a department. Our customers benefit, because enquiries are dealt with much more quickly, and directly handled by a specialist, addressing the right courses of action. With this structure, QM processes are also much closer to our personnel, so that quality runs through at every level of Tecan’s operations. It encourages very open communication, allowing everyone concerned to work together effectively.
Complicated flow charts have been replaced with easy-to-follow diagrams and pictorial ‘process arrows’, with keywords from the corresponding regulations, which take minutes rather than hours to understand. The new structure is designed to give customers greater confidence in Tecan’s quality processes, by being able to see the organizational structure clearly. It simplifies working together with customers’ quality departments during audits by saving time that would
otherwise be spent on understanding our processes, and focus on actual customer projects – spending the time to discuss the real issues. The structure still adheres strictly to medical devices laws, but where the pharmaceutical industry (eg. 21 CFR 211) dictates that complaints, as well as final product release of each batch, must be reviewed by a quality assurance department, the corresponding QSR for the medical device industry (21 CFR 820) is
General Manager
Head of Quality Management
Head of Sales & Management
Head of Services & Consumables
Head of Research & Development
Product Improvement
Complaint Handling
Quality Engineering
Head of Logistics & Production
Technical Quality
Quality Assurance
Excerpt of the organization chart for Tecan Austria, showing the business segment structure
Tecan Journal 2/2008
Quality Control
CAPA-Handling
Document Control
to receive future copies of the Tecan Journal you’ll need to re-register your details at:
Q UA L IT Y
www.tecan.com/get-journal
Management process
Resources & support
Customer need regulatory BOI supplier
Solution to customer need
Product Life Cycle
In & In
Regulatory
Compass (Order Processing) M1
M5
M8
M10
Innovative product portfolio
Continuous improvement
Process overview, based on ISO 9001:2000
more flexible, only specifying a formally designated person or unit. This makes sense, not least because pharma law applies to batch numbers for hundreds or thousands of tablets which could directly influence human health for example, but batch size for device manufacturers like Tecan is just one; one device with one serial number. “At Tecan Austria, we have replaced the matrix structure with a business segment structure. I now report to the head of business operations, with a dotted line to the vice president of global QA,” explained Robert Reingruber-Breitwimmer, head of QM at Tecan Austria. “We began in 2006 by adding a number of quality engineering specialists to the R&D team, which helps us to take quality requirements into consideration in the early stages of product development. At the same time we reorganized the procedures regarding complaint handling. Eight technical experts, who already know the devices intimately, work in our expert line in Tecan Austria to cover the 17 products produced here; three of them are now trained as quality specialists, and they ensure that their colleagues follow the correct QA procedures.” “We implemented other improvements in 2007. The previous system, where the quality officers were in the QM department, had many connecting points between quality and logistics, production and incoming inspection. With
each interface, we have lost time and information, so it made sense for these quality personnel to work within L&P and are therefore much more in touch with what is going on, while still keeping a dotted line to me, the head of quality management. Similarly, we have also installed a new function called ‘technical quality’ within production, where four specially trained people are responsible for design transfer, and they give the head of L&P a quality status report, which is essential to prevent future problems in production.” The Qualität und Zuverlässigkeit (Quality and Reliability) article explained that, for a more efficient quality assurance system, the head of quality management should not be responsible for direct provision of product quality, and instead should have a role that is closer to the general manager and assist him to improve or develop new strategies and processes. Robert explained: “Several rules have been defined regarding the responsibilities of the Business Unit Head and me, which ensure smooth operation as well as safeguarding against changing management positions. The structure has worked well at Tecan Austria, and it has been well received by customers and personnel alike.”
Tecan Journal 2/2008
27
28
EV E NTS 20 0 8
Meet Tecan at these events in 2008 Americas ISBER Annual Meeting
Bethesda, MD
18-21 May 2008
LRIG Mid-Atlantic Technology Exhibition
East Brunswick, NJ
22 May 2008
American Association for Clinical Chemistry Annual Meeting
Washington, DC
27 Jul 2008
National Clinical Diagnostic Laboratory Conference
Jinan, China
7-12 Jun 2008
International BioExpo
Tokyo, Japan
2-4 Jul 2008
Advances in Microarray Technology
Barcelona, Spain
7-8 May 2008
IQPC Compound Management & Integrity
London, UK
20-22 May 2008
Het Instrument
Utrecht, Netherlands
20-23 May 2008
Glycan Forum
Berlin, Germany
23-24 May 2008
ESHG European Human Genetics Conference
Barcelona, Spain
31 May-3 Jun 2008
Forum Labo & Biotech
Paris, France
3-6 Jun 2008
International Congress of Genetics
Berlin, Germany
12-17 Jul 2008
Annual Microarray and Gene Expression Data Society Meeting
Riva del Garda, Italy
1-5 Sep 2008
DGTI Annual Congress of the German Society for Transfusion Medicine and Immunohematology
Düsseldorf, Germany
16-19 Sep 2008
Biotech Forum and Scanlab
Copenhagen, Denmark
23-25 Sep 2008
Asia and Pacific
Europe, Middle East and Africa
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, Karin Schneider 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.
For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative. This brochure may contain reference to applications and products which are not available in all markets. Please check with your local sales representative. 8 Plus 1 Access, Freedom EVOlution, Genesis FE500, GENios, HID EVOlution, HS 400, HydroFlex, LaserCheck, LS Reloaded, Magellan, MultiChannel Arm, NanoQuant Plate, PosID, PowerScanner, quad4 monochromators, Te-Care, Te-PoolSafe, Te-PS and Te-VacS are trademarks and Cavro, Freedom EVO, Freedom EVOlyzer, 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. © 2008 Tecan Trading AG, Switzerland, all rights reserved.
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 www.tecan.com Tecan Austria +43 62 46 89 33 Belgium +32 15 42 13 19 China +86 10 586 95 936 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 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 +41 44 922 81 25 REMP Switzerland +41 31 770 70 70 Japan +81 44 542 70 21 USA +1 508 429 2200
Tecan Journal 2/2008