Automated sample prep – the reality of proteomics in the clinic
Pages 6-9
Novel methods guiding cancer treatment
Pages 10-11
Improved prostate screening to help men live long and prosper
Pages 12-13
Taking new cancer cures from bench to bedside
Pages 14-15
Cancer remains a growing global healthcare issue, and precision medicine is revolutionizing research, diagnostics and treatment in oncology. A major driver for this progress is the rapid advancement in genome sequencing technologies, which continue to reveal genomic mutations that can be targeted by tailored therapies. A growing body of knowledge is helping clinicians to further understand diagnostic markers and disease pathways underlying many cancers, leading to new methods for earlier diagnosis and advanced, personalized treatments.
State-of-the-art sequencing instruments are required to perform the complex assays that are fundamental to the development of precision oncology, and sequencing laboratories are calling for innovative technologies that can meet the rising throughput demands and aid the interpretation of vast quantities of data. This is essential to fully understand cancer mechanisms and, ultimately, develop more effective and timely treatments to improve patient outcomes.
The automation of sequencing workflows for higher throughput and greater reliability is undoubtedly increasing, allowing libraries to be generated overnight – with no requirement for manual input – ready for analysis the next morning.
Tecan has been a pioneer and market leader in laboratory automation for more than 40 years and offers a comprehensive and expanding range of innovative products and solutions. These technologies can be used to automate every stage of the NGS workflow, from liquid biopsy and tissue extraction to the preparation of RNA and DNA-based NGS libraries for sequencing in research and clinical settings.
With the acquisition of Paramit Corporation in 2021, we are ideally placed to expand our portfolio of complementary products for research, diagnostics and the medical field, now truly enabling us to scale oncology innovations from life sciences research to the clinic. Together, we offer a global development, production and support infrastructure that provides unparalleled support to our customers, and we contribute to better management of oncology and other diseases from basic research and early prevention all the way to late-stage treatment.
The translation of insights from technologies like sequencing into the clinical space will only increase in the coming years, and Tecan is excited to play a significant role in turning scientific breakthroughs in cancer research into practical healthcare solutions.
Achim von Leoprechting CEO
We may well be on the threshold of a new hope for oncology. Shorthanded to ctDNA, circulating cell free tumor DNA is sloughed off from tumors. It can be detected in liquid biopsies of just a few milliliters of blood. This could revolutionize what oncology can achieve, by diagnosing cancers earlier and more efficiently.
As a past presenter at the Molecular Diagnostics conference, Dr Theresa Zhang, Vice President of Research Services at Personal Genome Diagnostics, contends that ctDNA analysis is poised to have a profound impact on how we treat cancer patients in terms of diagnostics, prognostics and drug monitoring. In the lab, automation will be key to achieving both cost-effective throughput and the control over contamination that is critical to find the needle in the haystack: a few mutant DNA copies shed from a tumor into the blood and surrounded by thousands, or even tens of thousands, of normal copies. We
took the opportunity to ask her about the current status and future promise of ctDNA analysis in oncology.
Theresa is very enthusiastic about the potential for ctDNA in transforming oncology in practice: “Measuring ctDNA will help us with earlier detection in the future. It will be invaluable in studying minimal residual disease and reducing the level of over-treatment. For many cancers, diagnosis is followed by surgery, and then the tumor tissue is analyzed to decide on the risk of recurrence and the need for chemotherapy. That kind of risk assessment hasn’t been very accurate, and the result is over- or under-treatment. For example, in early stage breast cancer, many patients receive chemotherapy, even though large-scale clinical trials have shown that only 10-15 percent of patients really benefit from chemotherapy.”
“ctDNA tests can be helpful in identifying those patients. ctDNA tests done on blood draws a few weeks after surgery can be used to determine
whether the patient has residual tumor cells that are shedding ctDNA into the blood. If there is ctDNA in the plasma of patients after surgery, those patients will have a high likelihood of recurrence. The ability to detect early signs of recurrence could change the way we do risk assessment. Instead of statistical prediction, we are actually detecting recurrence.”
“Another aspect of diagnostics is directing targeted therapies in latestage patients. For example, patients who have specific mutations in the epidermal growth factor receptor (EGFR) are eligible for specific treatment with EGFR inhibitors. A plasma-based version of the EGFR assay can be used as a diagnostic tool in some patients. Two of these assays, marketed by Roche and Qiagen, have already been approved in Europe.”
Using ctDNA to monitor response to drug treatment is another hot area, as Theresa said: “Changes in circulating tumor DNA during treatment are very informative of response to treatment.
The analysis of ctDNA, which is released from any lesion in the body, often detects response to treatment or relapse from treatment earlier than a conventional approach, such as imaging, which is directed at only a few lesions. Increased levels of ctDNA can be detected months before imaging can detect that the tumor has returned. It’s very exciting that we are able to detect a molecular relapse so early; this means that we know when the therapy has stopped working long before the imaging assay indicates there is a problem.”
“Being able to do a test non-invasively – and get real-time information about the tumor – is also valuable, because the tumor evolves during treatment or as the disease progresses. What we have from the diagnostic block – the archival tissue – is not informative enough to guide treatment decisions later on. Right now, we are limited to archival samples, and real-time rebiopsies are not an option in most clinical settings. So ctDNA analysis offers an excellent alternative to serial re-biopsy, especially for late-stage cancer patients, since their tumors tend to shed more ctDNA into blood, which makes it easier to detect.”
The pros outweigh the cons –reliability is high Theresa acknowledges that there may be some disadvantages with ctDNA analysis, but they are definitely outweighed by the advantages: “With the current detection approach, not all the tumors shed detectable levels of ctDNA. And if the detection method is not sensitive enough, you will probably miss patients. But the biological background is very low for ctDNA – it only comes from cancer. The dogma is that, if you find ctDNA in plasma, then it means that you have cancer. We now know that some white blood cells may mutate clonally and shed mutated DNA into blood as we age, but these can be easily identified.”
After 12 years of experience at Merck leading a molecular profiling group supporting oncology, Theresa joined Personal Genome Diagnostics two years ago, just when the ctDNA field was really starting to pick up. The company analyzes ctDNA and interrogates sequence mutations, copy number changes and translocations in a large panel of 63 clinically-actionable genes. Detection is based on NGS, and the company is aiming at CLIA approval in the near future.
“Plasma Select-R has already been used by many pharmaceutical companies for retrospective analysis of their clinical trials samples. It helps them to understand the genetic landscape of the patients enrolled in their clinical trials, which is often uncharacterized, since patients have gone through many lines of treatment. Previously, pharmaceutical companies have had to rely on archival tumor blocks taken years prior to the trial, and were missing the real-time status of the tumor. So, where possible, they are now using liquid biopsies to assess the molecular landscape of the cancer they are treating.”
“Analysis of ctDNA also gives insights into possible mechanisms of drug resistance by the tumor. By comparing pre-treatment, baseline ctDNA results with the blood samples taken throughout the treatment, it is possible to determine if tumors have acquired new mutations, and if these new mutations have contributed to resistance.”
Theresa has a clear view on the need for automation in the clinical laboratory when it comes to analyzing ctDNA:
“Automation is key whenever a highly complex assay – such as an NGS-based ctDNA assay – is involved, because the assays are too complex to be scaled up
in a reliable and controlled manner. Moreover, automation is critical for highly sensitive ctDNA tests. Indeed, we need to detect one mutant molecule out of 1,000 and, in the future, out of a background of 10,000 wild type molecules. So contamination becomes a real problem. While automation is certainly cost effective, it’s also the key to controlling contamination.”
Automation of protein sample preparation workflows to be able to address clinical questions has come of age: it is both possible and realistic, despite the hurdles. Here, we discuss recent developments in reproducible, high throughput proteomics sample preparation workflows that do not require modification, even when applied to highly disparate sample types. These advances bring proteomics a significant step closer to clinical reality.
Not another ordinary attempt at ‘simplifying proteomics’
There are four major bottlenecks to being able to bring proteomics techniques to the clinic: the ability to analyze enough samples (hundreds to thousands or more), with a simplified workflow that is both time- and cost-efficient. Below, we will describe how new technological developments differ to previous claims and products, and support the bold assertion that they truly bring proteomics in clinical settings closer to reality. While tests developed with such workflows might require FDA regulatory approval, many CLIA LDTs (laboratory derived tests)
using the instruments and workflows described here are already in use, and we anticipate that robust automation will ease, and even promote, the development of many new LDTs.
Automated protein sample preparation must faithfully recapitulate the underlying proteome, regardless of the sample type. Further, interfering substances incompatible with downstream processing – like viral transport media (VTM) detergents, salts, glycerol, PEG, Laemmli loading buffer and biological contaminants
(eg. bile components) – must be fully removed. This step likely requires a solid support for capturing proteins.
While non-solid support solutions for automation exist, including bead- and liquid-based systems, they can require extensive optimization when working with complex biological samples, and often have issues with changes of scale. Ultimately, these limitations add complexity, time and cost. In the end, the stability of a solid phase support system remains unrivaled.
Filter-aided sample preparation (FASP) used to be frequently employed as a membrane support to remove sodium dodecyl sulfate (SDS) prior to mass spectrometry analysis.1 However, FASP protocols are notoriously long, and membranes fail regularly, leading to experimental failure. Such disadvantages led to the development of new solid phase capture strategies. In particular, the widely adopted S-Trap™ technology (ProtiFi) allows the use of high concentrations of SDS in a fraction of the time of FASP, with proteins captured on a true solid phase surface.
Independent research compared the yields from in-solution, FASP, and S-Trap based digestions of proteins extracted in SDS and urea-based lysis buffers. 2 Label-free quantification was performed to analyze the differences in the identified proteome using each method. The results showed that, while
the different digestion methods were reproducible within the method type, S-Trap outperformed FASP and in-solution digestions by yielding the most efficient digestion, with the greatest number of unique protein identifications
Uniquely, the S-Trap gives the ability to explore the insoluble portion of the proteome, even from such tough substrates as bone or muscle, by first fully solubilizing the samples and then converting the formerly insoluble proteins into soluble peptides. The product enables the extraction, solubilization and handling of all proteins in high concentrations of SDS, prior to further denaturation via reduction and alkylation, acidification (pH <1) and exposure to high concentrations of organic solvent. This four-stage denaturation ensures complete destruction of undesired enzymatic activity – such as proteases and phosphatases – and maximizes the efficiency of downstream digestions. Reduction and alkylation are performed in five percent SDS, precluding precipitation, or can be performed on-column.
Denatured proteins are captured, concentrated and rapidly cleaned of contaminants in the submicron pores of the S-Trap. Proteins are then digested in situ in a rapid (<1 hour), reactor-type digestion. Capture of protein, washing (SDS and contaminant removal) and
protease addition requires less than five minutes. After a one- or two-hour digest at 47 °C, peptides are eluted and ready for downstream processing.
S-Trap sample processing technology is available either as spin columns, or automatable 96-well plates. Both formats enable the use of SDS in proteomics sample preparation. S-Trap technology has been used for samples as diverse as serum and dirt and, more recently, has been extensively used to study SARS-CoV-2 infection in COVID-19 research.
Increasing throughput and efficiency: positive thinking
Simplifying the protein sample prep workflow requires a robust protocol that can automate the protein capture, cleaning and digestion steps. The ideal protocol should be identical for the highly varied sample types of interest, because sample preparation is the largest source of experimental variation – up to 75 percent according to published studies. 3
Complex protein samples can have heterogeneous and viscous consistencies, so automation with more robust positive pressure is preferable: vacuum at a maximum of one atmosphere may not be enough, and centrifugation requires expensive and complex mechanisms. Positive pressure has been shown to be more reproducible than vacuum, 4 and
typically operates at a higher pressure than vacuum approaches. Positive pressure can therefore more reliably process all the columns of a parallel set-up of a matrix of samples, such as in a 96-well plate, with constant pressure over a specified period with a defined pressure profile.
A realistic assessment of the requirements of a simplified, automated
protein sample prep workflow should include processing captured protein samples using a positive pressure unit with liquid dispensing capabilities, for cleaning and digesting samples prior to analysis. S-Traps are available in a 96-well plate format that is compatible with multiple automated platforms, including the Resolvex® A200 positive pressure workstation.
S-Trap sample preparation in combination with the compact, benchtop Resolvex A200 workstation offers affordable, accessible and high throughput automated proteomics sample preparation to laboratories for the first time. The Resolvex A200 system uses gas-based positive pressure to deliver maximum process reproducibility and uniformity across columns or wells, and automates accurate liquid dispensing for up to 11 protocol solvents, including the S-Trap denaturation, washing, binding and elution buffers. This ensures efficient clean-up, digestion and elution. The standard S-Trap proteomics protocols come preinstalled on the Resolvex A200 workstation, or users can create custom protocols optimized for unique needs, reducing processing times and enhancing analytical performance.
How far can S-Trap take us in the clinic?
Automation of the proteomics sample preparation will make a huge difference to the clinic in many key areas. One such area is in the mining of the millions of archival FFPE (formalin-fixed, paraffin-embedded) samples held in labs around the world. S-Trap has already been successfully used in this area, and the standardized protocol includes the use of the HYPERsol protocol, 5 which has been extended to the world’s only simultaneous 96-well megasonicator, the PIXUL™ (Active Motif). The HYPERsol protocol affords the best correlation ever published
between paired FFPE and flash frozen samples, as determined by both proteomics identifications and quantifications.
Having solved the bottlenecks associated with sample prep – being at last able to use the same, scalable protocol from sub-microgram to multiple milligram sample sizes, and having obtained new and standardized -omics data with the implementation of these simplified workflows – the next challenge is what to do with the data.
Automated sample processing enables generation of large numbers of samples, which brings with it a new problem: how do we make sense of all the data? The sheer quantity of data – along with the myriad tools and approaches – can lead to significant errors in data analysis, due to faulty assumptions and tools, or analyses being incorrectly applied. Indeed, this problem will only grow with the increasing sample throughput afforded by automation.
An efficient solution to this problem might be an online, cloud-based and browser-accessible -omics analysis engine, which translates data into biological meanings through an easy, accessible and interactive interface. ProtiFi recently developed such software, a world first. Called SimpliFi™, it accepts quantified -omics data of all kinds as input, obligates QC of the data,
and then performs many statistical analyses to allow users of all skill levels to explore and visualize their data. The interactive analyses include differentially expressed pathways, GO enrichments, cellular markers, proteinprotein interaction networks and molecular-level classifications, all of which can be performed on a smart phone. Results can be shared or published simply by sending a URL. SimpliFi uses all nonparametric statistics and resampling to provide accurate results and confidence intervals, regardless of the structure of the underlying data.
So, can we automate protein sample prep… or not? To revisit the original set of challenges in bringing proteomics into a clinical research setting:
Can we analyze enough samples, and can we simplify our proteomics workflow, both time-efficiently, and cost-effectively?
Yes, we can. Tecan and ProtiFi have partnered to create an integrated, highly scalable and easy-to-use proteomics workflow powered by the S-Trap sample preparation system and the Resolvex A200 automated positive pressure workstation.
Can we then analyze and understand the data?
Yes, SimpliFi allows non-omics experts to use these powerful tools to interrogate and understand biological questions.
1) Wiśniewski, JR et al. Universal sample preparation method for proteome analysis. Nature Methods, 2009, 6(5), 359-362.
2) Ludwig, KR et al. Comparison of in-solution, FASP, and S-Trap based digestion methods for bottom-up proteomic studies. Journal of Proteome Research, 2018, 17(7), 2480-2490.
3) Piehowski, PD et al. Sources of technical variability in quantitative LC-MS proteomics: human brain tissue sample analysis. Journal of Proteome Research, 2013, 12(5), 2128-2137.
4) https://www.americanlaboratory. com/914-Application-Notes/172423Evaluation-of-an-Automated-SolidPhase-Extraction-Method-UsingPositive-Pressure
5) Dylan, M et al. HYPERsol: High-quality data from archival FFPE tissue for clinical proteomics. Journal of Proteome Research, 2020, 19 (2), 973-983.
For research use only. Not for use in clinical diagnostics.
To find out more about Tecan’s positive pressure sample preparation solutions, visit diagnostics.tecan.com/samplepreparation-resolvex-positivepressure-processors
For more information about S-Trap technology and SimpliFi, visit protifi.com
Multiple myeloma (MM) remains an incurable blood cancer, but recent advancements in treatment have significantly improved patient prognosis and quality of life. UK-based specialist diagnostic company Binding Site has developed a new strategy that monitors pre-cancerous conditions associated with MM, as well as treatment success, helping to guide therapeutic pathways with more accurate information.
MM has a worldwide incidence of around 160,000 cases per year,1 and is more predominant in the elderly. It is unknown what causes MM, but a condition called monoclonal gammopathy of unknown significance (MGUS) is a common precursor, and requires constant monitoring to ensure early diagnosis if it progresses to malignancy. Traditional methods to diagnose MGUS involve measuring the monoclonal immunoglobulin M-protein through serum protein electrophoresis or immunofixation electrophoresis, which are also used to gauge treatment response in MM patients. These techniques have long been considered the gold standards, however, with the introduction of new therapies, patients are getting a better
depth of response, making it harder to monitor treatment success and identify relapses in a timely manner.
Binding Site focuses on medical solutions to this kind of challenge –particularly for blood cancers, such as MM, and immune system disorders –and has developed a more accurate method that will help with diagnosis and treatment monitoring for these patients. Stephen Harding, Chief Scientific Officer at Binding Site, explained: “MM patients usually present with a high disease burden, with somewhere in the region of 10 9 malignant cells in their bone marrow. Electrophoresis gels are sensitive to a concentration of around 107.5 , and so can accurately identify these patients
on initial presentation. The problem arises after therapy, where the number of malignant cells remaining – termed the minimal residual disease, an important prognostic factor revealing treatment success – is much lower, making it difficult to quantify cells accurately. Furthermore, antibodybased therapeutics can skew electrophoresis results, as therapeutic monoclonal antibodies (tmAbs) can be wrongly identified as endogenous monoclonal proteins and interpreted as residual disease.”
“We have developed a method using MALDI-TOF mass spectrometry that measures serum M-proteins and has significantly improved the sensitivity of quantification, meaning we can now monitor patients with much lower levels of the protein. However, sample preparation is extremely intricate, involving the extraction of immunoglobulins from patient samples at high volumes, and spotting onto MALDI plates at low volumes. Samples are pipetted from primary tubes and incubated with beads, which are then washed to clear all the serum proteins, ready for spotting. This last step often involves the transfer of liquid down to a single microliter, requiring incredible precision to deliver accurate results.”
Knowing the challenges associated with setting up an automated lab workflow, the company decided to look for an OEM partner to speed up the process, ultimately choosing Tecan Synergence™ to help with the project. Stephen continued: “We approached various vendors with these requirements, and decided to partner with Tecan because of the company’s experience with automation. We chose the Fluent® 780 Automation Workstation, and worked closely with Tecan from the beginning to refine and improve our workflow.
For example, we needed greater accuracy in some of the pipetting steps – the spotting especially – and a stronger magnet to pull down the beads, as we wanted to wash the samples vigorously. Each time we went to Tecan with specific requirements, the team always managed to find a solution, allowing us to achieve the desired precision and throughput. The entire workflow has been reduced to just two manual steps – loading the primary tubes and transferring the MALDI plates onto the mass spectrometer – making it an incredibly efficient process.”
“From there, mass spectrometry is able to deliver extremely accurate results, with a sensitivity of about 15 milligrams per liter for monoclonal proteins, compared to 1,000 to 3,000 milligrams per liter with gel electrophoresis. It also offers two-dimensional separation, allowing us to see novel features of monoclonal proteins – including post-translational modifications – which can correlate to different disease pathologies. The technology itself is easy to use, and we’re continuously working on intelligent software to remove the subjectivity of analyzing results, making it even more accurate and predictable compared to manually reading electrophoresis gels. This will prove increasingly important as more tmAbs are developed and routinely used, further contaminating the spectra and complicating analysis.”
“The partnership with Tecan has been vital during the development of this method. The Fluent is such a fantastic workhorse, and it really decreases the required hands-on time for staff. The
biggest differentiator, however, has been the nature of the partnership. The Tecan team often didn’t have an off-the-shelf solution but, as they understood our goals exactly, they always came up with a tailored approach. Without this support, we truly believe we wouldn’t be launching this product this year, and it also gives us the confidence to continue this collaboration. In the future, we might move to a fully integrated model –including the mass spectrometer – to increase throughput, or continue to refine the method to improve its accuracy even more.”
References
1) Ludwig H, Novis Durie S, et al. Multiple Myeloma Incidence and Mortality Around the Globe; Interrelations Between Health Access and Quality, Economic Resources, and Patient Empowerment. Oncologist 2020; 25(9):e1406-e1413. doi:10.1634/ theoncologist.2020-0141
This product is under development and not commercially available. Its future availability cannot be ensured.
To find out more about Tecan Synergence OEM systems engineering services, visit partnering.tecan.com/synergence
To find out more about Tecan’s Fluent Automation Workstation, go to www.tecan.com/fluent
For more information about Binding Site, visit www.bindingsite.com.
The Fluent is such a fantastic workhorse, and it really decreases the hands-on time for staff. The biggest differentiator, however, has been the nature of the partnership.
Prostate cancer is the second most common cancer in men, killing hundreds of thousands each year. However, the current routine screening method for checking prostate-specific antigen (PSA) levels generates false positives in an alarming number of cases, leading to thousands of unnecessary biopsies each year. To combat this, Life Length has developed ProsTAV ®, a state-of-the-art in vitro diagnostic test that uses telomere-associated variables to identify patients with a higher risk of prostate cancer, aiding screening initiatives and reducing the number of unnecessary biopsies.
The current standard of care used worldwide to screen patients for prostate cancer risk is by measuring their PSA levels, referring individuals with high concentrations for a biopsy. This procedure is not only invasive, painful and costly, it is often unnecessary. In fact, over two thirds of men who undergo a biopsy do not actually have cancer, highlighting the
need for complementary tools to more accurately predict the risk of prostate cancer before biopsy. The use of telomeres as predictive and prognostic markers for oncological diseases is a rapidly expanding research area. Life Length is an innovative biotech company that was spun out of the Spanish National Cancer Research Center in 2010, and has since gone on
to become a world leader in telomere diagnostics and measurement. Its biomarker technologies are now being used in everything from biotechnology and pharma to cosmetics and veterinary applications.
Life Length was part of the Horizon 2020 funded ONCOCHECK project to clinically validate the use of telomere-associated variables (TAVs) as biomarkers for different cancer types. The project was particularly successful at identifying lung and prostate cancers, and the results from 1,200 biopsied patients allowed the company to develop highly precise and predictive algorithms to determine a patient’s risk of prostate cancer. This approach has since been developed into ProsTAV, a state-of-the-art in vitro diagnostic test to identify patients with an elevated risk of suffering from aggressive prostate cancer.
The company uses its proprietary algorithm and machine learning techniques to analyze a combination of the patient’s age, PSA level, prostate exam results and TAVs, generating a prostate cancer risk score between zero and 100. The patient’s urologist can then use this score, in combination with other factors, to decide whether to refer the individual for biopsy. Stephen Matlin, CEO of Life Length, commented: “The objective of ProsTAV is to reduce
the number of unnecessary biopsies in men with elevated PSA levels. PSA testing leads to almost two million prostate biopsies a year, of which around 1.4 million end up testing negative. This highlights how inefficient PSA testing is; it frequently leads to extremely unpleasant biopsy procedures being performed on men that do not actually need it. Our objective with ProsTAV is to eliminate a significant number of these biopsies by replacing them with a combination of tests, including our sensitive telomerebased method. In fact, we recently received approval for the use of ProsTAV from AEMPS – the Spanish equivalent of the FDA – underlining the effectiveness of our approach.”
Life Length has recently invested in a number of Tecan laboratory automation solutions to improve its workflow and increase efficiency. “We prepare virtually all of our assays using our Fluent® and Freedom EVO® platforms, and the precision and flexibility of these solutions allow us to automate even our most complex tasks,” Stephen explained. “For example, our workflow includes a series of washes to remove cellular membranes while keeping the nuclei of the cells intact. It is a very delicate process to avoid damaging the
chromosomes, while removing everything else, but our Tecan instruments make it possible to do this in a completely automated fashion. The 384-channel pipetting head of the Fluent platform’s Multiple Channel Arm™ delivers superior repeatability over manual and semi-automated methods, while the Freedom EVOware® Sample Tracking software provides complete traceability of every sample prepared on the Freedom EVO platform. This combination of repeatability and traceability is essential in a diagnostic laboratory like ours, ensuring that samples are accurately processed and linked back to the correct patient.”
The success of the initial instruments has led to Life Length recently taking delivery of another pair of Fluent and Freedom EVO platforms to increase its output further. Stephen continued: “These additional workstations will significantly increase our efficiency; we can now run a set of tests on one pair of platforms while we write scripts and prepare the next set of tests on the second pair. This will not only maximize the use of our technicians’ time, but will also allow us to run more samples in a day, increasing our throughput.”
“The decision to purchase Tecan liquid handling systems was simple, thanks to
Tecan’s unrivaled support and service infrastructure in Spain,” Stephen added. “It was critical for us to have a local support network as, if a platform develops a problem, we need to be able to get support and source replacement parts quickly, without causing unnecessary delays to patients, who are waiting for extremely important – and potentially lifechanging – test results. It simply wasn’t an option to purchase solutions from other OEMs who don’t have a strong local presence.”
Stephen concluded: “With ProsTAV, we’re seeking to reduce the number of prostate biopsies by more than a third, eliminating unnecessary procedures. This will generate a significant clinical benefit for patients, and also provide substantial economic savings for both public and private healthcare systems. Now that our workflow has the precision and flexibility afforded by our Tecan platforms, we are aiming to become an essential service for urologists wanting to improve their diagnostic procedures.”
For research use only. Not for use in clinical diagnostics.
To find out more about Tecan’s liquid handling solutions for clinical applications, visit diagnostics.tecan.com/products/ liquid_handling_and_robotics
For more information about Life Length and its service, visit lifelength.com
We are aiming to become an essential service for urologists wanting to improve their diagnostic procedures.
Many of the anti-cancer drugs currently used for chemotherapy work by causing replicationassociated DNA damage that kills individual cancer cells. While this can be an effective way of treating the disease, the drugs often also indiscriminately affect healthy cells, causing unpleasant side effects for the patient. To help resolve this problem, researchers at the Karolinska Institute in Sweden are developing DNA repair inhibitors that are able to selectively introduce toxic DNA damage to cancer cells, while avoiding causing harm to normal cells, to support the successful treatment of cancers and improving patients’ experience of chemotherapy.
The Helleday Lab sits within the Department of Oncology-Pathology at the Karolinska Institute, and primarily investigates basic DNA repair, DNA damage signaling pathways and nucleotide metabolism. The lab’s overarching aim is to develop novel targeted therapies for cancer, inflammation and autoimmunity, as well as viral infections. The multidisciplinary team was among the first to demonstrate the synthetic lethality concept for the treatment of cancer, exploiting the high level of DNA damage and dysfunctional redox status present in cancer cells to prevent the repair of lesions and stimulate further injury to the cell. The advantage of this
approach is that these treatments should have far fewer negative side effects on patients than existing chemotherapies.
Dr Oliver Mortusewicz is the Principal Researcher and Basic Science Team Leader of the Thomas Helleday Research Team within the lab. He described the crucial work carried out by his group: “Our goal is to turn cancer defects into cures by targeting altered DNA repair and cellular metabolism found in cancerous cells. We aim to take our projects all the way from basic science to the clinic, and we have already been successful, with two Phase I clinical trials currently being
conducted using our MTH1 inhibitor candidate – karonudib/OXC-101 – at Karolinska University Hospital.”
The lab performs a wide range of tests as part of its drug discovery and development workflow, including target engagement assays to ensure the candidate molecule is binding to the target in the cancer cell, functional assays to test for the efficacy of drugs that induce DNA damage, and synergy assays to investigate how its potential anti-cancer compounds work synergistically with approved drugs. For all of these assays, the team needs to accurately dispense microliter, nanoliter, or even picoliter volumes of both candidate and approved therapeutics into 384-well plates prior to seeding the plates with cancer cells.
This low volume dispensing is difficult to do by hand accurately at the scale
The Thomas Helleday Research Teamrequired, and so the lab relies on a D300e Digital Dispenser to provide non-contact dispensing down to 11 picoliters. Before acquiring the D300e platform, staff had to perform these complex and extensive pipetting tasks manually, making them extremely time consuming, tedious and prone to human error. Oliver explained the difference the instrument has made to the lab’s productivity: “The D300e makes our lives so much easier, improving our efficiency and throughput by speeding up our liquid handling workflow and reducing hands-on time. In turn, this frees up the staff to do other vital tasks in the lab. Another significant benefit of the D300e is that it reduces the possibility of errors by automating many of the pipetting steps, which enhances our assay reproducibility.”
Oliver continued: “We also conduct resazurin assays to test for cell viability, but this test depends on the metabolic activity of the cells, which doesn’t correlate 100 % with cell viability. That’s why we also subsequently image the plates with a Spark® Cyto imaging plate reader. This image-based analysis allows us to get a better understanding of what is actually happening to the cells through the use of fluorescently-tagged proteins or immunofluorescence. For example, we can gauge the rate of proliferation or the localization of repair proteins upon the induction of DNA damage. A major advantage of the Spark Cyto over a classical plate reader is its ability to perform real-time imaging and cytometry. For example, we can carry out automated stain-free cell counting and confluence assessment of a whole microplate in just one image per well using the system’s bright field imaging technology.”
Oliver and his colleagues now rely on the two Tecan platforms for many of
Researchers are using DNA repair inhibitors to selectively introduce toxic DNA damage to cancer cells
their workflows, and are looking forward to employing them in further vital research going forward: “Both of the platforms are so easy to learn, and the interface is very user friendly. We have many students working in the lab with us and it doesn’t take long to train them up on it. That means they can get started on experiments sooner, which is great for our assay productivity. In addition, we have discovered that one of our anti-cancer molecules is actually also active in preventing inflammation, so we are now venturing out into other areas of research with our drugs, following wherever the science leads us. We certainly plan to keep using our versatile Tecan platforms in the future to support us in our upcoming investigations,” Oliver concluded.
For research use only. Not for use in clinical diagnostics.
For more information about the Tecan D300e Digital Dispenser, visit tecan.com/d300e
To find out more about the Tecan Spark Cyto, visit lifesciences. tecan.com/plate-reader-live-cellimaging-cytometry
To discover more about the Thomas Helleday Research Team’s work, go to ki.se/en/onkpat/ research-team-thomas-helleday
Our goal is to turn cancer defects into cures by targeting altered DNA repair and cellular metabolism found in cancerous cells.
Tecan Gives Back is an employee-driven initiative supporting charities that help to improve people’s health and quality of life. Created as part of the company’s 40th anniversary celebrations, the scheme gives employees the opportunity to earn ‘kilometers’ for their chosen charity by engaging in fitness and wellness activities, or by volunteering. These are then pooled and converted into cash donations at the end of the month-long challenge. One of this year’s beneficiaries is World Cancer Research Fund (WCRF), a global research charity focused on reducing the risk of developing cancer and improving the outcomes for those already diagnosed.
the risk of developing cancer, and supporting them in their pursuit of a healthier lifestyle. We also help individuals living with a cancer diagnosis, giving advice on nutrition and other factors that can improve their wellbeing and increase their chance of survival.”
Tecan’s mission is to add value to society by providing laboratory products that enable healthcare innovations, ultimately improving people’s lives and health. The ‘Tecan Gives Back’ initiative was created in 2020 as a part of this vision, and aims to promote a healthy lifestyle among employees while funding charities that support vital medical research. These goals are perfectly reflected in WCRF’s mission to help people to live longer, happier and healthier lives, free from the devastating effects of cancer.
Debbie Goddard, Head of Fundraising Development at WCRF, explained: “WCRF is the leading global cancer prevention charity, offering educational programs that teach people about how diet, weight and physical activity affect
WCRF’s educational programs and advice are grounded in decades of scientific literature, and the charity maintains close ties with the global research community. Its unique approach not only helps to fund vital cancer prevention research through direct grants, but also provides a holistic overview of current scientific understanding across a wide range of cancer-related topics. Dr Helen Croker, Head of Research Interpretation at WCRF, elaborated: “We have two main programs, our conventional Research Grant Programme – which funds novel cancer prevention and survivorship studies – and our flagship Global Cancer Update Programme. The latter provides an in-depth review of ongoing research, systematically assessing newly published peer-reviewed papers that investigate the relationship
between lifestyle and cancer risk, as well as those looking at factors affecting survival rates and health after a cancer diagnosis. We carefully examine not just the conclusions of those studies, but also the quality of the research performed, and use this data from across numerous sources to create an overview of advances in understanding in this field. The main advantage of having these two programs is that we can identify the gaps in the current knowledge, and work to fill them by funding certain research projects.”
“When we started our mission in the 1980s, the connections between cancer and parameters such as diet, weight and physical activity were controversial,” Helen added. “WCRF has since made a huge difference to global policy and advice by firmly establishing these as modifiable risk factors for cancer. We also look at the underlying mechanisms behind our findings, helping to understand how certain factors relate to cancer at a biological level.”
Debbie continued: “Studies have shown that our cancer prevention recommendations are very effective in reducing the probability of various cancers, as well as other diseases with similar risk factors, such as diabetes and heart conditions. This type of education can have a huge impact on public health, as there are approximately 375,000 new cancer cases every year in the UK alone and, according to our research, up to
40 percent of those cases could have been avoided. The funds received from Tecan will help us continue our research and spread awareness about cancer prevention that can ultimately help to save lives.”
Martin Brändle, Senior Vice President of Corporate Communications and Investor Relations at Tecan, added: “We are delighted to be supporting WCRF and its research efforts into new cancer prevention strategies
through the Tecan Gives Back initiative. This important work –alongside the efforts of other forward-thinking medical research and healthcare charities – fits with our vision of scaling healthcare innovation globally, as well as our aim to drive future developments in oncology.”
For more information about World Cancer Research Fund, visit www.wcrf.org
The funds received from Tecan will help us continue our research and spread awareness about cancer prevention that can ultimately help to save lives.
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, Benjamin Peltier
Project Lead: Tecan Trading AG, Itziar De Julian Garate
Editor: kdm/UK www.kdm-communications.com
Editor: UP THERE, EVERYWHERE/Sweden upthereeverywhere.com
Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf, Switzerland, hello@tecan.com, www.tecan.com
Images that are not owned by Tecan have been reproduced with permission, and may have been taken prior to implementation of social distancing measures. To register for the Tecan Journal please go to https://www.tecan.com/tecan-journal © 2023 Tecan Trading AG, Switzerland, all rights reserved.
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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
