Tecan Journal Edition 03/2014 by Tecan Group Ltd.

TJ Edition 3/2014

Tecan Journal

Life Sciences and Diagnostics

Confidence in drug discovery pages 10 - 11

Easy PCR set-up with TouchTools™ PCR Wizard pages 16 - 17

Fit for King’s pages 20 - 21

An antioxidant assay for the real world pages 22 - 23

Welcome to the latest issue of the Tecan Journal

It’s been a busy summer here at Tecan, with the launch of the Fluent™ workstation generating a lot of interest from new and existing customers alike. This advanced laboratory automation solution has been on display at several major exhibitions around Europe and Asia, and we’re pleased to say that it has been very well received, with the first customer orders being readied for shipment as we go to press. More recently, the addition of IBL International – a market leader in microplate-based immunoassays – to the Tecan Group has generated a lot of excitement (see pages 6-7). This acquisition is an important step for Tecan, and is an excellent fit with our long-term strategic aims of helping to simplify laboratory workflows by offering fully integrated solutions – including hardware, software and reagents. Mass spectrometry-based diagnostics is another area that is benefitting from this approach, and this issue of the Tecan Journal also has news of a collaborative project – working in partnership with RECIPE Chemicals and Instruments GmbH – to offer validated solutions for fully automated mass spectrometry sample preparation. A number of laboratories in this fast-growing area of clinical diagnostics are also taking advantage of our recently launched AC Extraction Plate™ to simplify their workflows, and we have details of how King’s College Hospital, London, is using this innovative consumable device for antipsychotic drug monitoring. As always, this Tecan Journal also contains a variety of additional stories demonstrating the diverse applications of Tecan instruments around the world, from food science in Russia to biobanking in the USA. Enjoy the issue, David Martyr, CEO

CEO WELCOME TECAN JOURNAL 3/2014

pages 6 - 7

IBL International is a perfect strategic fit with Tecan’s long tradition of serving the clinical market

CEO Welcome

4 Revolutionary transfer tool enables automated Nested LiHa tip and tray handling without a gripper pages 14 - 15

pages 16 - 17

Researchers at ISPRA have been trialing the TouchTools PCR Wizard for the Freedom EVO® platform to simplify complex genetic testing workflows

5 Infinite® 200 PRO and Gas Control Module used to investigate cancer therapies 5

Fluent Laboratory Automation Solution tours European exhibitions

6 - 7

IBL International joins the Tecan family

8 - 9

Educating tomorrow’s scientists

10 - 11

Confidence in drug discovery

12 - 13

Feeding the Russian biotech market

14 - 15

Incorporating genomic medicine into cancer research and primary care

16 - 17

Easy PCR set-up with TouchTools™ PCR Wizard

18 - 19

Making transfusion even safer with blood genotyping

20 - 21 Fit for King’s 22 - 23

An antioxidant assay for the real world

24 - 25

Imagination knows no boundaries

26 - 27

A RECIPE for success

Leading the debate

28 Events

pages 26 - 27

CONTENTS TECAN JOURNAL 3/2014

Revolutionary transfer tool enables automated Nested LiHa tip and tray handling without a gripper The Disposable Transfer Tool is an innovative new consumable device offering increased throughput and capacity for Freedom EVO® workstations. This patent pending tool is designed for use with Tecan’s Nested LiHa disposable tips, and offers fully automated handling of empty tip trays without the need for a gripper. One of the challenges that laboratories face when configuring their automated liquid handling systems is the need to balance capacity and functionality against space and budget constraints. Nested LiHa disposable tips offer increased worktable capacity for tip storage, allowing five trays of 350 μl tips to be stacked on a single SLAS-format carrier position, extending the walkaway times and improving productivity. Until now, this solution has only been available for instruments equipped with a Robotic Manipulator Arm or a MultiChannel Arm™ gripper option to remove empty tip trays. The Disposable Transfer Tool is a cost-effective

solution to overcome this, allowing the Freedom EVO’s Liquid Handling (LiHa) or Air LiHa Arm to pick up and dispose of empty trays. When the last tip is used, the LiHa simply fetches a Disposable Transfer Tool from the dedicated holder and uses it to transfer the empty tray to the waste. Implementation of the Disposable Transfer Tool is designed to be as easy as possible, with quick set-up and selection using Freedom EVOware® (v2.6 SP1 onwards). The only additional hardware required is a 16-position Transfer Tool Holder, which fits into a three-position trough carrier and can

The Disposable Transfer Tool picks up an empty tip tray and transfers it into the waste chute

PRODUCT NEWS TECAN JOURNAL 3/2014

be quickly and easily filled by hand prior to beginning a series of runs. This elegant solution is particularly suited to small or mid-sized Freedom EVO workstations – where workdeck space is limited – boosting capacity without a significant capital investment. It also offers benefits for larger systems, allowing the gripper to perform other operations while the LiHa Arm disposes of empty trays, improving productivity and functionality for high throughput applications. To find out more about Tecan’s Disposable Transfer Tool, visit www.tecan.com/consumables

Infinite® 200 PRO and Gas Control Module used to investigate cancer therapies Researchers in Salzburg, Austria, have been using an Infinite 200 PRO multimode reader and Gas Control Module (GCMTM) to study the efficacy of novel photodynamic therapies for skin cancers. Photodynamic therapies based on δ-aminolevulinic acid (ALA) potentially offer a highly selective approach for treatment of dermatological pre-cancers and early malignancies by causing a build up of the photosensitizing agent protoporphyrin IX in neoplastic cells. A collaborative team from

Paracelsus Medical University, the University of Salzburg and the Norwegian University of Science and Technology – along with representatives from Tecan – has recently published a study entitled Real-time analysis of endogenous protoporphyrin IX fluorescence from δ-aminolevulinic acid and its derivatives reveals distinct time- and dose-dependent characteristics in vitro1. This study takes advantage of the in-reader incubation capabilities of the Infinite 200 PRO and GCM to investigate the characteristics of novel ALA derivatives which might improve the pro-drug’s cellular uptake. To find out more about Tecan’s Infinite 200 PRO and Gas Control Module, visit www.tecan.com/gcm

The GCM enables incubation of cell-based assays within the Infinite 200 PRO’s measurement chamber 1 Kiesslich et al. J Biomed Opt, 2014, 19(8), 85007

Fluent™ Laboratory Automation Solution tours European exhibitions Tecan has been showcasing the recently launched Fluent Laboratory Automation Solution at exhibitions across Europe, giving scientists the opportunity to see firsthand how this innovative system could help to streamline their workflows and take the complexity out of cell biology research. The latest addition to the Company’s world-class laboratory automation portfolio, the Fluent workstation brings together all the modules and devices required for automation of a wide range of cell-based assays in a single efficient and easy-to-use system, and has already been on display at ELRIG Drug Discovery in the UK, MipTec in Switzerland, and World of Technology & Science in the Netherlands. Visitors to these shows had the chance to discuss the potential benefits of this powerful solution with Tecan’s applications specialists, as well as to learn

about the Company’s wide range of advanced automation solutions for cell-based and biochemical assays.

To learn more about Tecan’s Fluent Laboratory Automation Solution, go to www.tecan.com/fluent

PRODUCT NEWS TECAN JOURNAL 3/2014

IBL International joins the Tecan family IBL International – a leading provider of microplate-based immunoassays – has recently become part of the Tecan Group. Offering one of the widest ranges of specialty diagnostic assays for use in research and clinical laboratories, IBL International is a perfect strategic fit with Tecan’s long tradition of serving the clinical market with advanced laboratory automation instruments optimized for immunoassay processing.

assays are usually created by universities or smaller diagnostic companies, who then approach us because we have the development experience, manufacturing capabilities and sales network to make the test commercially viable.”

IBL International's range of assay kits can be used manually or automated

Microplate-based immunodiagnostics is a growing area which offers immunoassay tests for a variety of medical specialties, as well as routine testing for low throughput facilities. As this market segment is generally outside the scope of large in vitro diagnostics companies, there is an increasing need for integrated instrument and reagent offerings which provide preconfigured, user-friendly automation. The addition of IBL International to the Tecan family marks an important step towards better serving this market by offering fully integrated solutions – including hardware, software and reagents – to simplify laboratory workflows. James O’Brien, Head of Clinical Diagnostics at Tecan, explained: “IBL International is a leading company in the field of microplate-based immunoassay diagnostics, particularly specialty testing. This is an area that Tecan knows very well, having provided automation

CORPORATE NEWS TECAN JOURNAL 3/2014

of ELISA testing to customers around the world through both our Life Sciences and Partnering Businesses for many years. A relationship between the companies was a very logical step – it’s an association we are very proud of.” Established in 1983 in Hamburg, Germany, IBL International develops and produces antibodies and immunoassays – including enzyme, radio-labeled, and luminescence-based assays – for research and routine diagnostics. The Company focuses on the development and distribution of diagnostic assays for rare and specialty indications, working with academic centers and commercial partners to identify opportunities for the commercialization of new assays at an early stage of biomarker development. Dr Jan Boesen, CEO of IBL International, commented: “Prototype

“We have the experience and facilities to develop and manufacture immunoassays in large quantities. This includes both the specialized assays that we are well known for, and routine tests for customers who do not have high throughput requirements. These customers have the option to perform these tests manually or automate them on an open platform, such as Tecan’s Freedom EVO® workstation. Our first contact with Tecan was through this automated approach, as several customers were using our microplate-based assays on Tecan workstations.” “This kind of automation is becoming increasingly commonplace in laboratories around the world. It is obviously both time- and labor-saving, as well as reducing the risk of human error, but there is a variety of reasons that laboratories are opting for microplate-based testing. For many facilities – both in the Western world

Each kit contains all the reagents required to perform the assay

and in developing countries – low sample numbers mean that the cost of a large, closed system is simply prohibitive. Despite an increasing trend towards centralization of diagnostic services in many countries, hospitals still want to maintain some in vitro testing capabilities for urgent testing. Open automation systems are also appealing to some larger facilities, giving them the option to run a small platform for specialist testing in parallel with their main, closed immunoassay analyzers. With these factors in mind, we were actively looking for partners to help us offer complete automated testing, as well as to expand our sales network and product portfolio. We looked at various automation providers, and Tecan was clearly the best strategic fit. We were lucky that Tecan was simultaneously looking to increase its involvement in the diagnostics sector, so a relationship between the companies seemed logical.” “As a specialty testing-oriented company, we are focused on accelerating the transfer of new biomarker tests from the research bench to the clinical environment. As a result, we

have close ties with many of the pioneers in immunoassay-based diagnostics, and several of our assays are based on cutting-edge research. An example of this is our amyloid-beta assays for Alzheimer’s disease. Biomarker-based testing for Alzheimer’s is a relatively recent development – which has not yet been adopted by mainstream diagnostics providers – and our assays have the potential to significantly improve the accuracy of disease diagnosis. This is another area where our business is a good fit with Tecan, providing opportunities for many researchers already using Tecan equipment to quickly and easily access IBL products.” Tecan CEO Dr David Martyr concluded: "We are delighted to welcome the IBL International team into the Tecan family. The combination of our expertise will allow us to offer research and clinical laboratories a wide range of specialized immunoassays – together with optimized automation – supported by our global sales and service capabilities.” To find out more about IBL International, visit www.ibl-international.com

CORPORATE NEWS TECAN JOURNAL 3/2014

Educating tomorrow’s scientists Tecan Austria’s long-term collaboration with the Salzburg University of Applied Sciences has resulted in a national prize for Bachelor student Doris Rieder from the Austrian Society for Biomedical Scientists, and takes another step in the right direction with a project aimed at school children, teaching them about a healthy lifestyle.

Tecan’s team of microplate reader and washer specialists based in Austria are keen to share their expertise and knowledge to help to educate technicians and researchers of the future, and have established several collaborations with local and international science institutes and universities. One such agreement is with the Salzburg University of Applied Sciences (Fachhochschule Salzburg) which approached Tecan several years ago in relation to the Biomedical Scientists course at the Institute. Dr Christian Oberdanner, Marketing Application Specialist for Detection at Tecan, explained: “Tecan is a well-known major player in the life science industry here in Salzburg, and has an excellent reputation in this region. Fachhochschule Salzburg first

approached us in 2009 to provide some guest lectures for the Lab Automation and Measurement Techniques module, which my colleague Dr Katrin Flatscher and I continue to do to this day. All the students work routinely with microplate readers in the lab and, with our thorough product understanding and experience, we are very well placed to teach them all they need to know.” FH-Prof Priv-Doz Dr Geja Oostingh, head of the Bachelor degree course for Biomedical Scientists, added: “I had previous experience of Tecan from my time at the University of Salzburg where, although we routinely used microplate readers from several companies, the Tecan readers were by far the most versatile ones, easy to adapt according to what experiment we were doing. Our collaboration with Tecan works very well; it is important for our students to learn about

From left to right: Geja Oostingh, Head of Biomedical Analytics, and Mag Dr Doris Walter, Managing Director, both from Fachhochschule Salzburg, and Dietmar Fischerlehner, Head of Research and Development at Tecan Austria

CORPORATE NEWS TECAN JOURNAL 3/2014

An Infinite 200 PRO loaded up for delivery to the Fachhochschule Salzburg

microplate readers, about what’s inside, how they function, and what kind of assays can be run on them.” More recently, the agreement has extended to some students spending their internships at Tecan to see a different side of their specialty. In 2011, student Doris Rieder spent 10 weeks at Tecan preparing for her Bachelor thesis. “I was very interested in choosing a theme for my thesis that was different from the usual medical topics. At first, I felt quite intimidated by the proficiency of everyone at Tecan but, from the first day, I felt part of the team, and really appreciated how friendly and helpful everyone was. They had also obviously given a lot of thought to the project prepared for me. I investigated path length correction for absorbance measurements, comparing a mathematical approach with the standard reference method on three different volumes and using three types of microplate. It was a very worthwhile project and earned me the Society for Biomedical Scientists in Austria prize – sponsored by Abbott Laboratories – for the best national Bachelor’s thesis. I won 200 Euros for this, and presented my work at the Society’s annual meeting.”

In 2013, Tecan donated an Infinite® 200 PRO reader to the faculty to give students the opportunity to train on a state-of-the-art instrument and learn to perform different experiments alongside the Tecan lectures. The new reader is based at the University’s Salzburg Hospital site, where it is used for teaching and some associated research projects. One of these is a school-based project, as Geja explained: “The aim of the project is to educate school children about a healthy lifestyle. We take blood samples before and after a series of teaching sessions to see if the lessons have a positive effect on their health status. We measure a panel of thirty different analytes, including cholesterol, glucose and hormones etc, some of which are analyzed on the plate reader using a multiplex assay. The support we have had from Tecan has been excellent; we are very pleased with this collaboration.” Christian concluded: “We are delighted to be supporting local science education and look forward to continuing our relationship with the Fachhochschule Salzburg.”

To learn more about Fachhochschule Salzburg, go to www.fh-salzburg.ac.at To find out more on Tecan’s Infinite 200 PRO, visit www.tecan.com/infinite200

“The support we have had from Tecan has been excellent; we are very pleased with this collaboration.”

Dietmar Fischerlehner demonstrating the Infinite 200 PRO to students and staff

CORPORATE NEWS TECAN JOURNAL 3/2014

Confidence in drug discovery

A Fluent™ Laboratory Automation Solution is providing exceptional flexibility for small molecule screening at IME ScreeningPort in Hamburg, Germany. This groundbreaking system offers straightforward automation of complex assays, enabling both in vitro and cell-based assays to be performed on a single, compact instrument.

IME ScreeningPort – part of the Fraunhofer Institute for Molecular Biology and Applied Ecology – offers a full range of drug discovery research activities to academic and public research institutes, helping to bridge the gap between fundamental research and the pharmaceutical industry. Partnering with laboratories across the globe, IME ScreeningPort provides industrial-scale assay development and small molecule screening services, using advanced laboratory automation and informatics systems to provide high quality, validated drug candidates which can be transferred directly into pre-clinical pharmaceutical development pipelines. As an industrially-focused research institute working across multiple life science disciplines, IME ScreeningPort requires highly flexible, user-friendly automation solutions to ensure the

necessary throughput and efficiency. Dr Philip Gribbon, Assistant Department Head of IME ScreeningPort, explained: "We have a completely open approach when it comes to therapeutic areas, so we usually have around 20 projects ongoing at any one time across a variety of indications, including oncology, neurodegenerative, cardiovascular and metabolic diseases, as well as a number of neglected parasitic diseases. Our major strength lies in target-based assays – both biochemical and cell-based – but we also perform a large number of phenotypic assays, so need the flexibility to quickly switch between various assay formats to meet the timetables and demands of each project." Automation is an essential part of IME ScreeningPort's laboratory workflow, and the increasing popularity of high content, multiplexed assays meant that the Company

Left to right: Philip Gribbon, Gesa Witt and Markus Wolf with the Fluent system

DRUG DISCOVERY TECAN JOURNAL 3/2014

was looking for a new liquid handling and automation workstation capable of running these applications. After initial project discussions with Tecan in late 2013, it was clear that the Fluent platform offered both the flexibility and precision required for the drug discovery projects at IME ScreeningPort. Philip continued: "Having worked with Tecan in several of my previous roles, I was aware of what the Company could offer in terms of automated solutions to run microplate-based assays. A partnership was also a very good fit for both parties, providing a unique opportunity for us to benefit from Tecan’s application expertise and latest generation of liquid handling instrumentation, and for the Company to test the performance of this innovative solution in a real-world setting.” Following a one week training course for two IME ScreeningPort researchers at Tecan's Männedorf headquarters, the Fluent platform was installed in March 2014, and was quickly brought into operation for cell-based assays. Liquid handling and automation scripts for many of the assays were established in a very short time frame, enabling a rapid start on instrument testing and validation. "We initially identified five assay groups that would be good candidates for running on the Fluent and, working hand-in-hand with the Tecan technical team, we were able to develop protocols very quickly. The Fluent workstation's fully-integrated design is ideal for our needs, allowing us to incorporate many of the auxiliary devices needed to perform our assays into a single, compact workstation. Our platform has an Infinite® M1000 PRO multimode reader and a HydroSpeed™ plate washer – both of which are extremely good instruments – as well as an incubator and a carousel to store labware and assay plates. This gives us the capacity and flexibility to quickly switch between, for example, a kinetic assay that only takes a

few hours and a cell-based assay with long incubation times that can take several days, helping to ensure project deadlines are met and the instrument is being used efficiently." One of the IME ScreeningPort assays now benefitting from complete automation on the Fluent platform is screening for anti-cancer agents. Using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega) and cell lines from the NCI-60 panel, IME ScreeningPort has developed a protocol that allows triplicate dose-response curves to be generated for up to 100 compounds against 20 different cell lines in a single, unattended run. Performed in 384-well culture plates, this workflow offers exceptional throughput, and uses Fluent's three, task-specific arms and the integrated Infinite M1000 PRO to provide precisely controlled liquid handling and analysis, ensuring consistent, high quality results. “The Fluent solution offers far greater flexibility than the high throughput screening systems we were used to, allowing us to precisely define how liquid transfers and other operations are scheduled and performed. Although this level of functionality requires you to devote time and effort to fully understanding the system's capabilities, the interface and software are very user-friendly, and you are amply rewarded for your investment. So far, we haven't found anything that we can't do, and we've already identified a second tranche of assays that we'll be transferring to the Fluent platform,” Philip concluded.

"So far, we haven't found anything that we can't do."

To learn more about IME ScreeningPort, go to www.screeningport.com To find out more on Tecan’s Fluent Laboratory Automation Solution, as well as details of other cell-based applications which have already been automated on this system, visit www.tecan.com/fluent DRUG DISCOVERY TECAN JOURNAL 3/2014

Feeding the Russian biotech market Russia has been a fast-growing region for biotechnology and life sciences in recent years, with food and agriculture being key focus areas. Here is a snapshot from laboratories looking at the contrasting roles of microbiology in food production, both employing Tecan equipment in their workflows.

The Soyuzsnab Group of Companies is the largest supplier of food ingredients in Russia, and its portfolio includes the manufacture of food products from raw materials, the synthesis of flavorings, and the development of novel biotechnologies for food production. The development of sourdoughs and protective cultures for the meat, dairy and agricultural industries is an area of particular importance to Soyuzsnab, and the Company continuously invests in developing its strain collection in order to meet the changing needs of its customers. Until 2011, Soyuzsnab’s Biotechnology Laboratory used extremely time-consuming and labor-intensive manual microbiological methods for strain maintenance and manipulation, before purchasing a Freedom EVO® 150 system to increase efficiency. The workstation is configured for rapid screening of micro-organisms of interest in a

96-well format, and includes a Pickolo™ module for colony picking, an Infinite® F200 PRO multimode microplate reader and an integrated incubator. In a single experiment, the team can screen 96 different strains for characteristics such as salt and acid resistance (important for the production of cheese and sausages), growth rates, acid formation, resistance to bile and alkalis (important for probiotic cultures) and the formation of aromatic compounds, as well as investigating suitable growth media. The increased throughput offered by the Freedom EVO platform means that the Biotechnology Laboratory is using the system for a number of other applications, such as investigating random mutagenesis as a way of improving cultures. In addition to its normal application of transferring colonies to microplate wells, the Pickolo

The Pickolo module offers fully automated colony picking, plate seeding and MALDI spotting for Soyuzsnab

AGRICULTURE TECAN JOURNAL 3/2014

module is also being used to spot colonies onto targets for MALDI-TOF analysis, allowing the team to simultaneously look at species identification and characterization in one experiment. For all these applications, the operator simply needs to load the worktable and identify the colonies of interest either by manual selection or using predefined profiles. The method then runs automatically, with data available to analyze just a few hours later. Pavel Vyushinsky, Biotechnology Engineer at Soyuzsnab, commented: “We are very satisfied with the speed of our Freedom EVO workstation, which has taken on the workload of three skilled employees. It has also given us the ability to carry out long-term analysis of the physiological characteristics of micro-organisms in a 96-well format, which was not feasible before we had the system.”

The All-Russian Plant Quarantine Center is also using a Freedom EVO workstation in combination with an Infinite F200 PRO, this time using a NanoQuant Plate™. Part of the Federal Service for Veterinary and Phytosanitary Surveillance, the Center is the national plant protection laboratory, and is responsible for providing training to plant health and quarantine specialists from around the world, as well as the development of many procedural guidelines for the detection and control of plant pests and pathogens. The Bacteriology and Molecular Methods Laboratory specializes in the detection and identification of quarantine pests, such as the causative agents of fire blight (Erwinia amylovora), brown bacterial potato wilt (Ralstonia solanacearum) and corn bacterial wilt (Pantoea stewartii subsp. stewartii), as well as performing genetic testing for other laboratories within the Center. More than 9,000 samples are investigated by the Laboratory every year from across Europe, Africa, North and South America, as well as from Russia, using immunofluorescence, enzyme immunoassays, cultural and morphologic examination, biochemical techniques and molecular genetics. With such a high workload, the Laboratory invested in a Freedom EVO workstation with a Te-MagS™ module to automate routine sample preparation and speed up the screening of crop samples. The system carries out nucleic acid purification for 48 plant samples in parallel using reagents from Russian manufacturers Syntol and Biokom, as well as performing pre-PCR processing for sequencing on an Applied Biosystems 3500 sequencer (Life Technologies). Automating the process in this way has freed up two full time researchers to perform other tasks, significantly reducing purification

The All-Russian Plant Quarantine Center’s Bacteriology and Molecular Methods Laboratory team

costs per sample. Konstantin Kornev, Head of the Bacteriology and Molecular Methods Laboratory, explained: “A big advantage of the Freedom EVO platform is that it has allowed us to avoid the PCR inhibition often caused by starch and soil impurities, which was not always possible with manual extraction. The combination of the Freedom EVO with our Syntol and Biokom reagents has also further reduced the cost of extraction per sample, which is important for us.” In 2013, the Laboratory also purchased an Infinite F200 PRO NanoQuant to allow them to quantify DNA in up to 16 purified samples at once, alleviating another potential bottleneck in the workflow and greatly improving sample throughput. The quality of DNA from this process is now so good that it can be used for various downstream PCR and sequencing activities, contributing

significantly to the Center’s groundbreaking work in the investigation of plant pests. Konstantin concluded: “We really like the speed of the workstation, its accuracy and the quality of the DNA produced.” To find out more about Tecan’s liquid handling solutions, visit www.tecan.com/applications To learn more about Soyuzsnab, go to www.ssnab.com For more information on the All-Russian Plant Quarantine Center, visit www.vniikr.ru

AGRICULTURE TECAN JOURNAL 3/2014

Incorporating genomic medicine into cancer research and primary care The Sanford Health BioBank in the USA is using a Freedom EVO® HSM workstation and ReliaPrep™ Large Volume HT gDNA Isolation System for reliable, automated extraction of DNA from blood for genomic medical research.

Sanford Health in Sioux Falls is one of the largest rural healthcare providers in the USA, serving local communities in North and South Dakota, Minnesota, Iowa and Nebraska. In addition, it supports ‘world clinics’ located elsewhere in the US, China, Israel and Ghana. Originally Sioux Valley Health, the facility was renamed Sanford Health in 2007 in recognition of a $400m donation from its benefactor, Denny Sanford. Four years later, following a further $100m contribution in memory of his mother Edith, the Edith Sanford Breast Cancer Foundation was established, focusing on the incorporation of genomic medicine into breast cancer research and care. The Sanford Health BioBank was set up to support both this initiative and Sanford Imagenetics, a new enterprise dedicated to incorporating genetics into internal medicine. The biobank began collecting blood and tissue samples, primarily from oncology patients, for use in future research. Dr Chun-Hung Chan, Director of the Sanford Health BioBank, explained: “Our goal is to make more use of genetics, moving towards personalized medicine, and one of the driving forces behind the biobank is to ensure samples are available when they are needed, rather than slowly collecting samples once a protocol is approved. Although the biobank is still in its infancy, we have started distributing samples, mostly to Sanford Health investigators; once

BIOBANKING TECAN JOURNAL 3/2014

Left to right: Chun-Hung Chan, Megan Hanson and Shaina Kollis with the Freedom EVO HSM

the repository is large enough, we hope to also supply other researchers investigating similar health problems.” Prior to purchasing the Freedom EVO HSM workstation, Dr Chan attended a variety of meetings to establish the different options available, seeking advice from other members of the biobanking community, and was introduced to the Tecan-Promega partnership. The combination of Tecan’s Freedom EVO

workstation with Promega’s HSM 2.0 Instrument and ReliaPrep Large Volume HT gDNA Isolation System was ideal for Sanford’s needs. Dr Chan continued: “We have used Promega products for a number of years, so I knew they were reliable. However, I had no previous experience of laboratory robotics – Tecan was new to me – and I was reliant on recommendations and references from other users. I looked at quite a few different options, and what really sold the Freedom EVO HSM

“Manual DNA extraction of up to 32 large volume samples could take the entire day; with the Freedom EVO HSM it is completed in about four hours.” workstation to me was the huge amount of flexibility it offered for sample processing. At the time, comparable systems were centrifugation-based, requiring equal numbers and volumes of samples, whereas the Freedom EVO HSM system offered the flexibility to extract from 1 to 32 samples of varying volumes in the same run. It also allowed extraction of high blood volumes – up to 10 ml – helping to ensure that we would not run out of a particular DNA sample. As many oncology patients are immunocompromised and have low white blood cell counts, isolating DNA from the largest possible starting whole blood volume is necessary to ensure that sufficient DNA is purified for long-term studies.” Predefined methods for processing multiple sample types using the Freedom EVO HSM system and the ReliaPrep Large Volume HT gDNA Isolation System chemistry, supported by Promega’s installation and training services, enable laboratories to rapidly implement routine operation of the instrument. Prior

to purchasing the Freedom EVO HSM workstation, all the biobank’s samples were processed manually. Automating the biobank’s protocols on the Freedom EVO HSM system enhanced speed and throughput, as well as consistency, enabling staff to spend their time performing other tasks. Dr Chan highlighted the benefits of switching from manual to automated processing: “The Promega system is designed for both manual processing and automation, enabling straightforward transfer of our protocols to the Freedom EVO HSM system. This offers several benefits, particularly improved consistency and speed of processing. Manual DNA extraction of up to 32 large volume samples could take the entire day; with the Freedom EVO HSM it is completed in about four hours, freeing staff to carry out other tasks. Setting up our blood and saliva protocols on the system is easy too, as the TouchTools™ touchscreen user interface visually guides users through each processing method, including starting a purification run, providing intuitive assay set-up.”

Customer support was another important factor in Sanford’s decision to purchase a Freedom EVO HSM workstation. “I looked into the history of the various companies supplying automated systems, as well as the level of customer support, and felt that Tecan had the edge. Tecan has a strong background in automation – a history I felt comfortable with – and was able to provide all the support I needed. Tecan and Promega have been very supportive, and the system has proved robust and reliable. After on-site validation, the Freedom EVO HSM came into routine operation in spring 2012 and has been so successful that we have since purchased a second system,” concluded Dr Chan. To find out more about Tecan’s genomic solutions, visit www.tecan.com/genomics To find out more about the Sanford Health BioBank, visit www.sanfordresearch.org/biobank

BIOBANKING TECAN JOURNAL 3/2014

Easy PCR set-up with TouchTools™ PCR Wizard Researchers at ISPRA have been trialing the TouchTools PCR Wizard for the Freedom EVO® PCR workstation to simplify complex genetic testing workflows. This software enables rapid selection and walkaway set-up of pre-PCR protocols suitable for a wide range of downstream analytical techniques.

The National Institute for Environmental Protection and Research (Istituto Superiore per la Protezione e la Ricerca Ambientale, ISPRA) is an Italian public organization performing technical, scientific and research projects related to environmental issues. ISPRA’s Laboratory of Genetics, based in Ozzano dell’Emilia in Bologna, investigates the genetic variability and composition of wildlife fauna in Italy, monitors the dynamics of threatened Italian mammals – such as the brown bear, wild cat, otter and wolf – and performs genetic

studies on endemic species including the Italian hare and Italian roe deer. The Laboratory also supports forensic investigations against poaching and illegal trade of endangered species on behalf of the Ministry of the Environment, including paternity testing of species listed in the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) to verify the legal origin of captive-bred animals. Most of the analytical methods used by the Laboratory – including phylogenetic studies, paternity testing, genetic monitoring and population genetics – are based on PCR techniques. Samples – including blood, feathers, hairs, buccal swabs, feces, urine and

tissues – are supplied to the Laboratory in standardized collection tubes which contain appropriate solutions for the optimum conservation of biological materials. This also ensures standardization of samples for the Laboratory’s automated protocols, speeding up the workflow and improving reliability. Dr Nadia Mucci, technologist, explained: “We purchased two Freedom EVO platforms in 2010 to automate our pre- and post-PCR sample processing. The first platform is used to automate DNA extraction in 96-well plates and set up PCR reactions in either 96- or 384-well plates. After pipetting, the plates are transferred to offline PCR thermocyclers for amplification. At the end of the PCR run, the plates are loaded onto a separate Freedom EVO platform for post-PCR processing, which prepares the samples for sequencing on our Applied Biosystems 3130xl genetic analyzers (Life Technologies).” “We chose Tecan instruments because we appreciated the technology, the support from Tecan’s application specialists, and the user-friendly software which offered us the opportunity to develop automated protocols for many more applications. The high throughput and wide range of downstream analysis methods used by our Laboratory mean that we need a highly reliable, very flexible solution, and the Freedom EVO certainly offers that.”

The ISPRA Genetics Team with Research Director Professor Ettore Randi

GENOMICS TECAN JOURNAL 3/2014

“To further improve our workflow, we recently tested the new TouchTools PCR Wizard with our broad range of DNA markers on a Freedom EVO 100 PCR workstation equipped with an Air LiHa air displacement pipetting arm. This software has proved extremely useful when we have a lot of different combinations to perform for our PCR experiments. For example, in forensic paternity testing, we usually test several related animals

using a species-specific panel of at least a dozen DNA markers. Because each study involves a different number of relatives, and we have around 150 different species-specific panels, this often required a new protocol using our existing platforms. This is no longer necessary with the TouchTools PCR Wizard, as we do not need to individually program every combination of markers and individuals.”

“We need a highly reliable, very flexible solution, and the Freedom EVO certainly offers that.”

“It is also very useful for inexperienced users, allowing them to set up and run protocols without the need for programming knowledge. Colleagues who rarely used our existing Freedom EVO workstations are now interested in working on the platform with the TouchTools PCR Wizard, and more experienced team members find it very useful for complicated PCR experiments. Moreover, processing is now much faster, thanks to the Air LiHa’s multi-dispense function and the seamless transfer of plate layout data to downstream instruments.” “I would like to thank ISPRA and Tecan for giving our Laboratory the opportunity to test the TouchTools PCR Wizard. It offers straightforward automation of PCR reaction set-up on Freedom EVO platforms and allows effortless operation, with clear visual representations and step-by-step instructions on the touchscreen. It guides the user through the entire process and simplifies sample preparation for a wide range of PCR-based techniques, such as sequencing, genotyping, DNA profiling, gene expression and detection of pathogens, reducing training time and costs,” Nadia concluded.

Federica Mattucci operating the post-PCR Freedom EVO system

To find out more on Tecan’s PCR solutions, visit www.tecan.com/pcr To learn more about ISPRA, go to www.isprambiente.gov.it

The TouchTools PCR Wizard visually guides the user through instrument set-up

Plate layout data can be easily exported for downstream instruments

GENOMICS TECAN JOURNAL 3/2014

Making transfusion even safer with blood genotyping AXO Science has established a Freedom EVO®-based high throughput blood genotyping system, helping to meet the needs of blood banks and transfusion centers around the world.

AXO Science, based in Lyon, France, specializes in the development of high throughput and multiplexed microarray diagnostic solutions for characterization of biological samples. With extended blood group genotyping now seen as the next step towards even safer blood transfusion, AXO Science has collaborated with the French National Blood Service (EFS) to establish a rapid, automated high throughput blood genotyping solution based on its patented HIFI Technology. Samuel Serraz, Director of Sales and Marketing, explained: “Blood banks in France needed a large-scale blood genotyping solution, but were struggling to find a system that was both high throughput and cost effective. We collaborated with EFS, using our technology to develop the HIFI Blood 96™ assay, establishing a robust, automated blood genotyping process on Tecan’s Freedom EVO workstations.” R&D Manager Benjamin Corgier took up the story: “HIFI Blood 96 is a multiplexed assay performed in a 96-well plate, and is fully automated on two Freedom EVO 100 platforms. DNA extraction is performed using a MACHEREY-NAGEL NucleoSpin® 96 Blood Kit – a silica membrane technology – on a Freedom EVO system equipped with a four-channel Air LiHa air displacement pipetting arm with adjustable tip spacing, a Robotic Manipulator (RoMa) Arm, a Te-VacS™ vacuum separation module, and an Eppendorf ThermoMixer®.” “Initially, 200 µl of whole blood is transferred to the lysis plate, and 40 µl of lysis buffer is added. After lysis, the samples are moved onto the filter plate, where the DNA is bound to the silica membrane, washed and finally eluted, producing around 80 µl

CLINICAL DIAGNOSTICS TECAN JOURNAL 3/2014

of extracted DNA. The plate is sealed and transferred to a thermocycler for DNA amplification, in preparation for the analysis. High throughput post-processing and hybridization is then carried out on the second Freedom EVO platform, which is equipped with a MultiChannel Arm™ 96, a RoMa, a HydroSpeed™ washer, a plate reader, three Pinnacle thermal heaters – two fixed at 37 °C, the other used to perform cooling temperature gradients – plate holders and tip racks.” “High throughput processing is based on hybridization of the amplification product on AXO Science microarrays printed in 96-well plates. The PCR product plates are placed on the workdeck and the amplified DNA is transferred to plates containing the microarrays. A cooling temperature gradient, from 90 to 60 °C, is then used to obtain specific hybridization on the microarrays. We characterize single nucleotide polymorphisms (SNPs), and can detect a single base difference in the amplicons. From this point onwards, the protocol is based on different wash steps, performed on the HydroSpeed, followed by incubation with labeling enzymes. Finally, incubation with the substrate results in precipitates forming at the different spot positions – each array has 49 spots – making them appear darker. Optical density measurements are then used to determine the signal, analyzing the images using the integrated AXOware software. We process 14 SNPs in duplicate, plus controls and dark spots for positioning the grid during detection and, because everything is automated, continuity and traceability are ensured from beginning to end.” The open architecture of the Freedom EVO workstation means that both the pre- and

post-PCR processing can be performed using the same platform and software, reducing the training requirements for new operators. Samuel continued: “We chose the Freedom EVO because it is a low maintenance system that we were already familiar with. Everything is controlled by Freedom EVOware® software, which launches the protocols and works with the AXOware image analysis software to create output files that allow us to analyze the results and create reports. HIFI Blood 96 received the CE IVD mark in January 2014, and we recommend the use of the Freedom EVO platform to automate our solutions.” “The Freedom EVO 100 is ideal for our purposes, and has allowed us to develop an automated, large-scale blood group genotyping process, meeting the need for

a rapid, high throughput protocol that is reliable and cost efficient. For customers at blood banks needing even higher throughput, there is the flexibility to choose one of the larger Freedom EVO platforms and, if demand is sufficient, we also have the capability to print onto 384-well plates, which would enhance throughput further still,” concluded Samuel.

“The Freedom EVO 100 is ideal for our purposes, and has allowed us to develop an automated, large-scale blood genotyping process.”

To find out more about Tecan’s clinical solutions, visit www.tecan.com/clinicaldiagnostics To find out more about AXO Science, visit www.axoscience.com Not all products discussed in this article are available in all countries. Contact your local Tecan Sales organization for more details.

Freedom EVO 1

Freedom EVO 2

The HIFI Blood 96 assay offers automated processing of 96 samples in just 4 hours and 20 minutes

CLINICAL DIAGNOSTICS TECAN JOURNAL 3/2014

Fit for King’s Tecan’s AC Extraction Plate™ and Freedom EVO® liquid handling platform have enabled scientists at King’s College Hospital to fully automate the analysis of clozapine and norclozapine in plasma, saving time and increasing sample throughput.

King’s College Hospital, London, is home to a large clinical pathology laboratory offering diagnostic testing services for both the hospital’s patients and external samples from across the UK and further afield. Managed by Viapath – a joint venture commercial partnership – the Laboratory has developed a number of automated sample preparation protocols to deal with growing sample numbers and the trend towards high throughput liquid chromatography-mass spectrometry (LC-MS) techniques for a variety of analytical

procedures. Lewis Couchman, Senior Clinical Scientist (Clinical Biochemistry Laboratory and Toxicology Department), explained: “One of our most frequently requested assays is for monitoring the drug clozapine – used in the treatment of schizophrenia – and we currently perform about 30,000 tests a year for this drug alone. At present, these samples are prepared manually by liquid-liquid extraction but, with numbers continuing to increase, we wanted to automate the analytical process. The idea was to make it more rapid, more robust and less prone to human error, as well as reducing costs and providing the capacity to grow the service in the future.” “Our department was a test site for the AC Extraction Plate for the analysis of 25-hydroxy-vitamin D in serum and, following this evaluation, I felt this technique could

work equally well for our clozapine assay. We carried out some proof-of-concept experiments with excellent results, and approached Tecan. The Company agreed to collaborate with us to further develop the application, and installed a Freedom EVO 100 workstation equipped with an eight-channel Liquid Handling Arm using disposable tips, a Te-Shake™, 16-position sample racks and space to accommodate 96-well sample extract plates, all controlled by Freedom EVOware® software.” Lewis continued: “Initially, we validated the AC Extraction Plate methodology using our existing calibration approach, with analysis of plasma blanks at the beginning and end of every plate, as well as calibration standards and QC material. However, we have now also shown that it generates equally good results

“Automating the AC Extraction Plate procedure on the Freedom EVO makes a real difference to our workflow.” Professor Bob Flanagan, Dr Caje Moniz and Lewis Couchman with the Freedom EVO workstation used for automated extraction

TOXICOLOGY TECAN JOURNAL 3/2014

“Controlling the Freedom EVO with Freedom EVOware is very easy; even though the software was new to us, it is really intuitive. With the AC Extraction Plate protocol established, we are fully flexible and can use the same workstation to prepare vitamin D and clozapine samples. There are potentially another 10 to 15 compounds that should work just as nicely using the AC Extraction Plate, including six or seven drugs used in psychiatry that are similar in structure to clozapine. We’re also looking at moving our steroid analysis onto the AC Extraction Plate, as the current sample preparation methodology takes a long time to perform.”

conjunction with a plate-based autosampler, eliminating the need for LC and only taking about 10 seconds to analyze each sample. We would then be able to transfer extracts directly from the AC Extraction Plate onto an LDTD plate, and analyze the entire plate in as little as 15 minutes. Our turnaround times would go from days, with manual processing, to just a few hours, which would be fantastic for our therapeutic drug monitoring service!” “We’ve had the Freedom EVO for just a couple of months, but huge amounts of data have already been generated. We have compared

To find out more about Tecan’s AC Extraction Plate, visit www.tecan.com/acplate To find out more about Viapath at King’s College Hospital, visit www.viapath.co.uk

Clozapine

1.80

Norclozapine

1.60 1.40

y = 0.9749x + 0.0031 R2 = 0.9897

1.20 1.00 0.80

y = 1.029x - 0.0347 R2 = 0.9394

0.60 0.40 0.20 0.00 0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

2.00

[Analyte] (mg/l) using manual extraction

Correlation between manual liquid-liquid extraction (x-axis) and automated extraction using the AC Extraction Plate (y-axis) Sample addition

Sample removal step

Analyte extraction

Sample Sample additionaddition Sample addition Sample addition

: analytes Sample Wash addition step : phospholipids

Analyte extraction Wash removal step

: salts : proteins

Wash

“Looking forward, we hope to further improve our throughput by evaluating laser diode thermal desorption ionization (LDTD) in

around 500 samples prepared manually by liquid-liquid extraction and using the automated method, and are really impressed with the results; there is no discrepancy between the two methods. Currently, we are in the final stages of the evaluation phase, but everybody is excited at the prospect of seeing it implemented,” concluded Lewis.

2.00

[Analyte] (mg/l) using automated extraction

using an isotopic internal calibration method. Instead of a batch calibration curve, we add a mixture of isotopically-labeled clozapine and norclozapine standards at three different concentrations covering our calibration range to each sample. This approach gives us an individual, matrix-matched, three-point calibration curve for each sample, and all compounds show excellent recovery. We just need to run QCs to check the validity of the result, and we can generate and report results straight away, without waiting for a batch to finish. It also increases our sample throughput from 65 samples per plate to between 80 and 85 samples, decreasing the cost per test. Automating the AC Extraction Plate procedure on the Freedom EVO makes a real difference to our workflow. Previously, samples arriving at the laboratory by 1 pm would be manually extracted during the afternoon and then run overnight, reporting the results the following day. The Freedom EVO prepares a whole AC Extraction Plate in around 30 minutes, and samples can be run immediately on our LC-MS, with a runtime of just under four minutes a sample.”

Sample removal Analyte elution

Wash removal

: analytes : analytes

: analytes : salts

: salts : phospholipids : :analytes : :salts phospholipids proteins

: phospholipids : phospholipids : proteins

Analyte elution

: salts : proteins

: proteins

The AC Extraction Plate provides a simple extraction protocol

TOXICOLOGY TECAN JOURNAL 3/2014

An antioxidant assay for the real world Researchers at CIRAD have developed a simple antioxidant assay allowing direct measurement of the antioxidant capacity of food samples. Based on UV absorption, this rapid technique is being performed on an Infinite M1000® PRO.

The French Agricultural Research Center for International Development (Centre de coopération internationale en recherche agronomique pour le développement, CIRAD) is a publically-owned industrial and commercial enterprise working with a range of developing countries to further scientific research and support international agricultural development. Its Performance of Tropical Production and Processing Systems Department centers its research on the interaction between biological processes and environmental conditions, providing advice and technical interventions to improve

agricultural practices. Within the Department, the Agropolymer Engineering and Emerging Technologies Research Unit (UMR IATE), based in Montpellier, uses a range of techniques to assess and improve crops and plant biomass derivatives, from increasing the nutritional content of whole plants or active ingredients to modifying the properties of macromolecules and composite foods. One area currently under investigation within the UMR IATE is the antioxidant capacity of foods. The presence and efficacy of antioxidants within foodstuffs is of ever-increasing interest to nutritionalists and consumers alike, yet the assays required to assess antioxidant capacity are generally laborious and ineffective, requiring organic solvents and offering a poor reflection of true in vivo activity. To overcome this, the Unit has developed a CAT (conjugated

Left to right: Mickaël Laguerre with PhD students Claudia Grajeda and Erika Zago

READERS TECAN JOURNAL 3/2014

autoxidizable triene) assay1 which allows routine screening of pure plant extracts for antioxidant activity. Mickaël Laguerre, the researcher who developed the CAT assay as his PhD project, explained: "The aim was to create a simple, relevant method that could be easily transferred to other labs or even used for routine measurements in industry. For this reason, we wanted to develop a high throughput assay using an aqueous medium instead of the organic solvent often used in this situation. We also wanted to employ a lipid-like substrate that would be more representative of in vivo activity." Mickaël and his colleagues developed a microemulsion-based optical method using commercially-available stripped tung oil as an oxidizable substrate. Triacylglycerols (TAGs) from the oil contain eleostearic acid which has conjugated triene groups (three

conjugated double bonds) that exhibit strong UV absorption at 273 nm. Under oxidizing conditions, degradation of conjugated triene TAGs leads to bleaching at 273 nm. This process can be delayed by the presence of antioxidants, enabling the quantification of the antioxidant activity in a sample. Mickaël commented: "A UV absorption method seemed to be the simplest option, but there were several significant obstacles to this approach. The first issue we encountered was that the pro-oxidative compound used to generate peroxyl radicals in the assay is extremely temperature sensitive, making it impossible to achieve consistent results in a standard cuvette. As a result, we required a microplate reader with very precise temperature regulation." "We looked at several options, and the Tecan instrument was the best fit for our needs, combining format flexibility and excellent temperature control with monochromator‑based optics. This set-up is ideally suited to assay development, as it allows complete freedom in wavelength selection, and also makes it more versatile for other research activities. The ability to evolve the instrument as your needs change – adding fluorescence, luminescence or polarization functions – is also a significant advantage, as your assay development choices are not limited by the reader." "The Infinite M1000 PRO is very user‑friendly, as is the Magellan™ data analysis software. One feature that I particularly like is the option to perform fully automated data analysis – providing a quick result for screening applications – or export of the data to an Excel® spreadsheet. It's probably true of all researchers, but I like to have the raw data available for more in-depth analysis, and it is very easy to do that with Magellan."

Erika Zago operating the Infinite M1000 PRO

As well as providing the reader, Tecan's application team was instrumental in translating the method from a research project to a routine assay. Mickaël continued: "When I first started, I was using a quartz microplate to allow measurements at 273 nm, which was both tedious to use and very expensive. The Tecan team put me in contact with Greiner Bio-One, as the company was about to launch the UVStar® microplate, and this was a real breakthrough in the development of the assay." "Since completion of the project, this assay is now in routine use, and we are also employing the Infinite M1000 PRO for a variety of other applications. We perform a number of ORAC assays and fluorescence‑based measurements, as well as UV absorbance measurements to follow, for example, the stability of polyphenols. Overall we are very happy with the system."

"This set-up is ideally suited to assay development, as it allows complete freedom in wavelength selection."

1Laguerre M, López Giraldo LJ, Lecomte J, Baréa

B, Cambon E, Tchobo PF, Barouh N, Villeneuve P. Conjugated autoxidizable triene (CAT) assay: A novel spectrophotometric method for determination of antioxidant capacity using triacylglycerol as ultraviolet probe. Anal Biochem 2008; 380:282-290.

To find out more on Tecan’s Infinite M1000 PRO, visit www.tecan.com/infinitem1000pro To learn more about CIRAD, go to www.cirad.fr

READERS TECAN JOURNAL 3/2014

Imagination knows no boundaries

AstraZeneca scientists engaged in surface plasmon resonance studies are reaping the benefits of a recently acquired HP D300 Digital Dispenser, saving time, enhancing consistency and using significantly less compound for dose-response experiments.

The Structure and Biophysics department at AstraZeneca R&D Mölndal, Sweden, is focused on the application of biophysical methods to determine how compounds interact with drug targets, examining physical parameters such as thermodynamics and binding kinetics. Principal Scientist Stefan Geschwindner explained: “Our department is involved in everything from determining the feasibility of a potential project to supporting lead identification, lead optimization and even characterization of candidate drugs for clinical studies. We aim to understand the driving forces behind compound-target interactions through a range of biophysical approaches, principally surface plasmon resonance (SPR) and nuclear magnetic resonance (NMR), but also technologies such as thermal shift assays and isothermal titration calorimetry.”

Stefan continued: “The need for the HP D300 Digital Dispenser was largely driven by a demand for more effective SPR studies and increased flexibility. We have set protocols for structure-activity relationship screening, using 7 and 10 concentration dose-response curves with a fixed range of dilution patterns and starting concentrations. However, there are times when we need the flexibility to use different dilution patterns to correctly describe dose-response curves and determine exact affinity values, for instance when cooperative binding is observed. Primary fragment screening is also a major part of our work; after dissolving the solid compounds in DMSO to prepare 100 mM stock solutions, manual dilutions are typically performed in 96- or 384-well plates, which is very time consuming. We needed to look at alternative solutions, as well as to widen our scope for the usual compound studies.”

Stefan Geschwindner setting up a dose-response curve on the HP D300

DRUG DISCOVERY TECAN JOURNAL 3/2014

“The HP D300 is very easy to use and saves us so much time. It gives us the flexibility to use very low concentration stock solutions and perform different dilution patterns, and the instrument does all the calculations for us. Another big advantage is that we save a lot of material, which is particularly important when the amount of compound available is limited; once the 10 mM stock solution has been prepared, there can be very little material remaining. Where we may only have sufficient compound to run one dose-response curve if acoustic dispensing were used, the HP D300 can even recover any solution that has not been dispensed, which is brilliant. The savings are tremendous and, other than manual pipetting, this is probably the only effective way to prepare our dose-response curves.” “It is also much easier to set up SPR experiments, which are very sensitive to variations in DMSO concentrations. Using the HP D300, we have found that, when compounds in DMSO are dispensed directly into a buffer without DMSO, the resulting DMSO content is so low that we can eliminate DMSO from the running buffer in many cases, particularly for very potent drug candidates. We now have a lot of data showing that this is a very straightforward way of preparing dose-response curves and,

without the need to make two buffers or run solvent correction cycles, we save lot of time.”

“The limit of the HP D300’s capabilities is basically the imagination of the users.”

“The HP D300 is also beneficial for thermal shift assays, where a dye is used to monitor protein unfolding. We prepare a matrix of different protein versus dye concentrations, and also test varying volumes. Once we have the optimal volume and ratio of protein to dye, we use the HP D300 to dispense a constant concentration of dye into a 384-well plate. This gives us consistency throughout the lead screening exercise.”

curves with the acoustic dispenser. Everybody says that it is probably the best investment we have made in the last five years. The costs-to-benefits ratio is just brilliant, and it can do so much. The limit of its capabilities is basically the imagination of the user,” concluded Stefan.

“We’ve had the HP D300 for about nine months now, and are really pleased with it. It’s so intuitive that hardly any training is necessary; even inexperienced users can become proficient in basic operation in as a little as five minutes. Originally, it was just intended for our group, but now staff from all departments are using the system, as it is so much easier than doing dose-response

To find out more about the HP D300 Digital Dispenser, visit www.tecan.com/digitaltitration To find out more about AstraZeneca R&D Mölndal, visit www.astrazenecamolndal. com/more-about-molndal.php

55 45

Binding signal (RU)

“We already had an acoustic dispenser, but this was not ideal. As there are peaks and troughs in our work, we needed an economical solution that could accommodate these changing demands, and the HP D300 was an attractive option. We contacted Tecan, who provided an instrument for us to try, and people immediately saw the benefits, realizing that the HP D300 could really change our way of working, helping to generate data more rapidly and shorten lead times.”

35 25 15 5 -5 -30

120

150

180

210

240

Time (s) Sensorgrams of a 22 concentration-response experiment prepared with the HP D300 showing the binding kinetics of a small molecule engaging with a protein-protein interaction target

DRUG DISCOVERY TECAN JOURNAL 3/2014

A RECIPE for success

Tecan and RECIPE have worked together to automate ClinMass® LC-MSMS Complete Kits* for the analysis of vitamin D and immunosuppressants on a Freedom EVO® platform, entering into a co-marketing agreement to help meet the demand for high throughput sample preparation for mass spectrometry. RECIPE Chemicals and Instruments GmbH, based in Munich, Germany, was established in 1982 and has grown to become one of the leading producers of HPLC and LC-MSMS in vitro diagnostics (IVD) kits. With clinical laboratories increasingly turning to high throughput LC-MSMS analysis, there was a need for more rapid sample preparation techniques. Recognizing this, RECIPE sought to automate sample preparation for mass spectrometry, initially focusing on its immunosuppressant and vitamin D kits, as these are particularly high throughput assays in the clinical laboratory. Therapeutic drug monitoring of immunosuppressants is crucial for transplant patients, helping to ensure the long-term viability of a transplanted organ, while the incidence of vitamin D deficiency, which is associated with many chronic conditions such as rickets in children and osteomalacia in adults, is on the increase. Dr Johannes Engl, R&D specialist for the vitamin D assay, explained: “We developed the ClinMass LC-MSMS Complete Kit for 25-OH vitamin D2/ D3 in response to strong demand from clinical laboratories which were increasingly moving from HPLC to LC-MSMS. The assay is quite straightforward, involving the addition of either serum or plasma samples to a 96-well

“The automated protocols are about three times faster than the manual protocol, which allows significantly greater sample throughput.” 26

microplate containing a precipitation reagent and control standards. After shaking and ambient temperature incubation, the samples are centrifuged and the supernatant analyzed by LC-MSMS. However, when performed manually, it is time consuming; automation was clearly the way forward.” Silvia Baecher, R&D specialist for the immunosuppressant assay, agreed. “Like vitamin D analysis, there has been an increase in the number of samples received by clinical laboratories for therapeutic drug monitoring of immunosuppressants. Throughput is important, but because you are monitoring treatment there is an even greater need to maintain chain of custody and process security than with vitamin D. The protocol for the immunosuppressant kit is almost identical to vitamin D – except that homogenized whole blood is analyzed rather than plasma or serum – and, with careful choice of mass transitions, cyclosporine A, tacrolimus, sirolimus and everolimus can be simultaneously monitored by LC-MSMS.” With such a clear need to move from manual to automated sample preparation, RECIPE chose a Freedom EVO equipped with an eight-channel Liquid Handling Arm. Johannes commented: “Transfer of our existing manual vitamin D and immunosuppressant assay protocols onto the Freedom EVO proved quite straightforward; minimal modifications were required. We received very good support from Tecan to help us become familiar with the system and discover its capabilities, learning to build our own protocols. Samples were analyzed both

LC-MS SAMPLE PREPARATION TECAN JOURNAL 3/2014

manually and using the automated procedures, and the results compared really well. The big advantage of automating the assays on the Freedom EVO is speed; the automated protocols are about three times faster than the manual procedures, which allows significantly greater sample throughput. We also benefit from enhanced reproducibility and sample security. The potential for manual errors is removed, and the option to include a barcode reader offers traceability, which is crucial for the clinical environment. In addition, scientific staff are freed to perform more technical tasks in the laboratory.” Flexibility was another key issue, as RECIPE plans to automate more of its assay kits. Johannes continued: “We know that flexibility is important to our customers, and the Freedom EVO offers many possibilities. It is a very good instrument and you can integrate modules – centrifuge, plate readers – for almost anything you want to do. It also offers a choice of disposable or fixed tips, and we evaluated both with our assay kits. The results were very good, with no sample carry-over detected.” Dr Stefan Plank, Head of PR at RECIPE, concluded: “After just a few months, the automation project has been so successful that we have entered into a co-marketing agreement with Tecan. Our expertise in developing LC-MSMS assay kits will now be complemented by Tecan’s years of experience in laboratory automation, allowing our customers to benefit from greater throughput, and improved reproducibility and process security.” To find out more about Tecan’s LC-MSMS sample preparation solutions, visit www.tecan.com/lcms To find out more about RECIPE, visit www.recipe.de * Not for sale in USA

Cornelia Kegele, Head of Group Branding and Marketing Communications

ClinCal® Calibrators and ClinChek® Controls offer reliable calibration and quality control

Leading the debate How a company is perceived as a brand is a sum of all its actions; not just its products or services, but also how it interacts with individual customers and the wider scientific community. Your brand influences how you connect with others, becoming an important ‘touch point’ to drive communication with researchers and organizations across your markets.

Chromatogram of an immunosuppressant analysis

Laboratories in the life science sector want to work with trusted partners who can help to ‘shape their labs’ by delivering the expertise and solutions they need. Scientists believe in results, proven data and their peer groups, and choose brands that reflect their own vision and goals. In order to build a brand effectively, customers need to be able to associate your company with values and a promise, which should be communicated in your own unique style. This differentiates you from your competitors, bringing additional value to the business and acting as a driver of future success. Emotionally engaging messaging and exciting, relevant content – with a good balance of brand and product marketing – make for a healthy communications mix. In the future, companies will have to work harder to get the attention of their customers, but there’s no shortcut to success. What kind of communication will make people stop and read? To stand out from the crowd, you need to nourish your brand by developing products and services that perform well, communicating your world in an interesting way, and letting your customers tell your story. We hope you enjoy the new look Tecan Journal! To let us know your thoughts, contact us at hello@tecan.com Find out more at www.facebook.com/tecantalk

Correlation between manual and automated preparation of vitamin D samples

TALK TO TECAN TECAN JOURNAL 3/2014

TJ Meet Tecan at these events Americas SOFT 2014

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26 – 29 Oct 2014

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7th National Congress of the Chinese Society of Blood Transfusion (CSBT)

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11 – 12 Nov 2014

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Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276 Design: OTM/London www.otmcreate.com Photography: Günter Bolzern/Zürich www.bolzern.tv Editor in Chief: Tecan Trading AG, Cornelia Kegele Project Lead: Tecan Trading AG, Cornelia Kegele/Antonietta Allocca Editor: kdm/UK www.kdm-communications.com Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, journal@tecan.com, www.tecan.com To register for the Tecan Journal please go to www.tecan.com/journal © 2014 Tecan Trading AG, Switzerland, all rights reserved.

EVENTS TECAN JOURNAL 3/2014

Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law. In general, the trademarks and designs referenced herein are trademarks, or registered trademarks, of Tecan Group Ltd., Mannedorf, Switzerland. A complete list may be found at www.tecan.com/trademarks. Product names and company names that are not contained in the list but are noted herein may be the trademarks of their respective owners. For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative. This journal may contain reference to applications and products which are not available in all markets. Please check with your local sales representative: www.tecan.com/contact