Life Sciences, Diagnostics and Partnering
Analyzing poor quality RNA: how low can you go?
Pages 14-15
High speed immunodiagnostics with multicolor imaging
Pages 16-17
Seeing is believing: harnessing the power of automation in single-cell genomics workflows
Pages 20-21
Diagnosing hormone-based disorders using saliva
Pages 24-25
Our understanding of cancers and their causes continues to progress every year and, as a result, oncology services are moving away from a generalized, one-size-fits-all approach to personalized strategies focused on more predictive, pre-emptive and preventative models. The benefits are clear, but as our knowledge and capabilities grow, so too does the daunting task of assessing, diagnosing and treating every patient as an individual.
The missing link is the empowerment of healthcare systems to scale their approaches to screening and diagnosis, treatment and surgical intervention and, eventually, prevention. This is where Tecan can help. By working with our customers to scale healthcare innovation, we play a decisive role in accelerating cancer research and the development of targeted drugs, novel diagnostics and advanced medical treatments. We offer innovative sample preparation, laboratory automation, medical workflow and digital solutions that propel enabling technologies from research use to global clinical deployment. With this in mind, we recently partnered with Nature Medicine to host a select group of industry leaders, top clinicians and prominent researchers from across key areas of oncology. The two-day symposium brought together many diverse thinkers and solutions providers with a collective interest in sharing knowledge, opinions and expertise in a way that will move us all collaboratively closer to the next great achievements in oncology. We plan to carry forward that momentum with another symposium in 2024, and invite you to join us on our journey to continuously create spaces for discussions, debates and breakthroughs.
While cancer is a focus area for us, this issue of the Tecan Journal also gives you the opportunity to read about some of the many other application areas where Tecan’s broad knowledge and capabilities are helping to scale up even the most complex workflows, contributing to our objective of improving peoples’ lives and health around the world.
Achim von Leoprechting CEO
Myasthenia gravis (MG) is an autoimmune disease affecting 14-20 of every 100,000 people in the US,1 and 1-9 in 100,000 people in Europe. 2 The sad truth is that most of those afflicted go undiagnosed. MG causes severe muscle weakness, and significantly decreases quality of life. Diagnosis can be difficult, but state-of-theart disease biomarkers and targeted assays are available to increase the likelihood that a patient with MG will be diagnosed early, and can be treated appropriately. So how can these biomarkers, and use of the correct assays, help clinicians to monitor therapeutic efficacy and support better treatment outcomes for their patients?
In MG, the body’s own immune system produces antibodies that attack the skeletal muscles responsible for breathing and various other movements. This leads to patients experiencing muscle weakness, and prolonged rest is needed to recover. There are effective treatments for MG, 3 but they are only effective if diagnosis is made early enough.
In most cases of MG, autoantibodies target acetylcholine receptors, which are essential for transmitting electrical impulses through muscle tissue. These electrical signals stimulate the muscles to contract. Acetylcholine receptor autoantibodies (ARAbs) produced in MG patients prevent acetylcholine from binding to its receptor, which blocks normal muscle contractions.
Around 85 percent of people with MG have ARAbs, while the remaining 15 percent have so-called ‘seronegative’ MG. Despite having clear clinical symptoms, these patients don’t have detectable autoantibodies against the acetylcholine receptor. About half of these seronegative patients will instead have autoantibodies against a protein called muscle-specific tyrosine kinase (MuSK). MuSK plays an essential role in connecting the acetylcholine receptor to the muscles, making it important in MG. Patients with MuSK autoantibodies (MuSK-Abs) typically have a more severe form of the disease, with more debilitating symptoms.
ARAb and MuSK account for 85-90 percent of MG diagnoses, meaning that – even with good markers – there is no way to diagnose 10 percent of sufferers. One emerging marker that
has shown diagnostic promise is low-density lipoprotein receptor-related protein 4 (LRP4). One study tested patients who were seronegative for ARAb and MuSK-Abs for the presence of antibodies against LRP4. They discovered that 12 out of the 13 patients tested were positive for LRP4, making this one of the most promising markers to help close the gap on people suffering with undiagnosed MG. 4
What makes myasthenia gravis diagnosis so complex?
ARAb and MuSK-Abs are the gold standard biomarkers for diagnosing MG. However, getting to that crucial early diagnosis is not simple. The first step is an assay for ARAbs, if MuSKAbs are not also measured right at the beginning. However, an ELISA for ARAbs is not sufficient, and is likely to yield a false positive result. To maximize the sensitivity and specificity of ARAb detection, a radioreceptor assay (RRA) is the preferred choice. The RRA is preferable because, unlike during an ELISA, the three-dimensional structure of the acetylcholine receptor is retained, which is essential for antibody recognition.
If the RRA test for ARAbs is positive, it aids in the diagnosis of MG. However, if the result is negative, it is important to consider the possibility of seronegative MG. To identify these cases, a follow-up test with a MuSK ELISA is necessary, as it can detect around 50 percent of these patients. Due to this reason, more and more laboratories directly involve the measurement of MuSK-Abs in the first line diagnostic test. The preferred test for this is a quantitative MuSK-Ab ELISA, which is highly accurate and reliable.
It also follows that if the test for MuSK is negative, then LRP4 could be the target. However, as there are no commercial tests currently available, laboratories currently have to develop in-house assays, which is now very time consuming under the In Vitro Diagnostic Regulation (IVDR). In such cases, it is probably best to collaborate with a commercial partner to get new tests registered under IVDR and commercially available as soon as possible.4
Patients with MuSK-Abs not only have a more severe form of the disease, but the therapy they receive also tends to be more aggressive, commonly involving rituximab immunosuppression to achieve B-cell depletion. In the treatment of MG, the target reduction of ARAbs is 50 percent, whereas the clinical goal of treatment in MuSK-positive patients is complete elimination of the MuSK-Abs. Calculating when this has been achieved requires a highly sensitive and quantitative assay, rather than relying on qualitative results. Additionally, since therapy may require some time to take effect, clinicians can carefully monitor the efficacy of MG treatment by using a sensitive, quantitative assay. They can detect increases or decreases in MuSK-Ab levels, and modify both treatments and therapeutic decision making as needed. Thankfully, both ARAb and MuSK-Ab tests are commercially available under IVDR, and there will be a reliable supply going forward.
1. Myasthenia Gravis Foundation of America.
2. The Portal for Rare Diseases and Orphan Drugs.
3. National Institute of Neurological Disorders and Stroke. National Institutes of Health. Myasthenia Gravis.
4. Pevzner A, et al. Anti-LRP4 autoantibodies in AChR- and MuSKantibody-negative myasthenia gravis. J Neurol. 2012, 259 (3), 427-35.
Soluble interleukin-2 receptor: a biomarker for immune activation
The quantification of soluble interleukin-2 receptors (sIL-2R) in serum or plasma in adults has become an extremely useful tool for clinicians to assess immune function in vivo for the investigation, management or outcome prediction of a broad spectrum of diseases. However, the measurement of sIL-2R is simple in comparison with the complex story behind this biomarker, which has become so central to our study of immune-mediated diseases.
Why measure sIL-2R levels?
Interleukin-2 (IL-2) is a key signaling molecule in the human immune system. It is a cytokine: one of a group of small, secreted proteins released by cells that have a specific effect on interaction and communication between cells. As such, IL-2 regulates the activities of the white blood cells that are responsible for immunity, forming part of the body’s natural response to infection and helping it to discriminate between foreign (‘non-self’) and ‘self’.1
IL-2 mediates its effects by binding to IL-2 receptors (IL-2R), which are expressed by lymphocytes. The major sources of IL-2 are activated CD4+ T cells and activated CD8+ T cells.1 The sIL-2R is secreted – or ‘shed’ – on T cell activation, meaning that elevated concentrations of sIL-2R are found in patients suffering from any one of an extensive range of conditions associated with an ongoing immune response, including sarcoidosis, multiple sclerosis, biliary cirrhosis,
chronic immune activation in common variable immunodeficiency (CVID), and hemophagocytic lymphohistiocytosis (HLH). 2-6
The convenient implication of this association is that sIL-2R could hypothetically be used as a generic biomarker to monitor/predict disease activity and treatment response for these diseases, and many others.7,8 However, in order to be certain that we are measuring something that is clinically relevant, we must delve a little deeper into how the IL-2/sIL-2R system functions.
IL-2 and sIL-2R in immune activation: unravelling the twisted plot
Binding of IL-2 to sIL-2R can either enhance or reduce immune response, depending on the target cell being involved in immunity or selftolerance. 9,10 The history of the sIL-2R – also called sCD25, sTAC and IL-2RA – goes back to 1985, when it was first described as being actively released by activated peripheral blood T cells via proteolytic cleavage of the cell surface IL-2R.11 In 1990, Rubin and colleagues were the first to demonstrate that, after in vitro activation, T lymphocytes enhanced cellular IL-2R expression and released sIL-2R(α). The study also showed that
– similarly to cellular IL-2R expression – the release of sIL-2R required de novo protein synthesis rather than cellular proliferation.12 However, despite this long-recognized association between immune activation and increased sIL-2R release under pathological conditions, the biological actions of this molecule are still far from understood. Several mechanisms of action, ranging from immune-inhibitory to immunostimulatory effects, have been proposed.
sIL-2R binds IL-2 efficiently and, based on in vitro experiments, it has been proposed that sIL-2R may limit activation and proliferation of T lymphocytes by sequestration of available IL-2.13 However, conflicting data has been reported.14 Alternatively, sIL-2R complexing with IL-2 may prolong IL-2 half-life, possibly enhancing the immune-stimulatory properties of IL-2, even by activation of low affinity dimeric IL-2R.15
It is possible that IL-2 can be presented to CD4+ T lymphocytes through sIL-2R, which then induces differentiation into regulatory T cells (Tregs) – rather than differentiation into T helper cells (Th1 or Th17 lymphocytes) – which can subsequently suppress immune activity.16 On the other hand, there are reports to support observations that sIL-2R may promote (auto)immune processes in association with enhanced Th17 generation, which involves sequestration of the IL-2 that normally inhibits early Th17 differentiation.17 Despite the obvious complexity and lack of agreement so far regarding the exact mechanism(s) of action of different configurations of sIL-2R, as well as their in vivo occurrence and final biological effects, the data currently available supports the role of sIL-2R in regulating IL-2dependent cell function.
An increased level of sIL-2R in the blood is considered to indicate an ongoing immune response, and this could theoretically be used to monitor a huge range of immune-mediated diseases. This is a good starting point, even if the generic nature of the response might initially be of concern. 9 Generally speaking, conditions that are characterized by excessive production of lymphocytes – so-called lymphoproliferative disorders – show high sIL-2R levels compared to healthy controls. This also applies to granulomatous diseases, such as sarcoidosis, in which T cell activation is a typical hallmark. The relative stability of sIL-2R levels throughout adult life, and the minimal gender-related differences, are further useful attributes that help make sIL-2R an attractive biomarker.12,18 That said, individual differences in age, gender, lifestyle and general health may need to be taken into account when considering reference values and ranges.19
sIL-2R levels are generally measured using immunoassays – either enzymelinked immunosorbent assay (ELISA), or chemiluminescent immunoassay (CLIA). There is very good correlation between the results of both assay types, although absolute values of ELISA (pg/ml) are about 7-8 times higher than those obtained by CLIA (U/ml). Commercially available ELISAs
should be calibrated against the international reference standard NIBSC 97/600.
ELISAs for sIL-2R are typically designed for batch analysis, and have a measuring range up to 10,00020,000 pg/ml, based on a standard serum dilution of ~1:5. Further dilutions will, of course, enable the quantification of higher sIL-2R levels. This process can be streamlined by the use of automated liquid handling systems, so that multiple dilutions can be analyzed simultaneously.
In clinical practice, laboratories will need to use different criteria for determining the upper limit of normal (cut-off) for sIL-2R levels; there is such a broad range of values that distinct cut-off values may be required for each situation. These will depend on the disease that is being diagnosed or monitored, and on the stage at which sIL-2R levels are measured for that disease.
There are many research use only (RUO) ELISA kits available for measuring sIL-2R levels. However, in order to be able to use a kit for diagnostic purposes, it must comply with applicable regulations. In the European Union, this means that kits used for IVD must conform to IVDR (Regulation (EU) 2017/746 for in vitro diagnostic medical devices).
The principal characteristics of sIL-2 kits might include, but are not limited to:
• A ll kit reagents and standards should be ready-to-use – no dilution necessary
• C alibrated against NIBSC 97/600 Standard Preparation
• I nternal kit controls provided
• E asily automatable
Sarcoidosis and sIL-2R: a deep dive Sarcoidosis, also known as BesnierBoeck-Schaumann disease, is a rare condition that causes small patches of swollen tissue – called granulomas – to develop in the organs. 24 It often affects the lungs and lymph nodes, and can also affect the skin. The symptoms of sarcoidosis depend on which organs are affected, but typically include tender bumps on the skin, shortness of breath and a persistent cough.
It is impossible to predict how sarcoidosis will affect a person, as the condition can affect any organ, and the symptoms vary widely depending on which organs are involved. Sarcoidosis may be acute or chronic, and some people do not have any symptoms at all, so that their condition might only be diagnosed after an X-ray carried out for another reason.
Elevated sIL-2R levels have been reported in numerous studies in sarcoidosis patients and, as such, sIL-2R is already an established biomarker for the disease. 2,25 Some studies suggest the measurement of sIL-2R as a marker of therapy success. 26 Sensitivity of serum sIL-2R
as a diagnostic biomarker for sarcoidosis lies around 79 percent and, in patients with uveitis, the sensitivity of elevated sIL-2R levels to establish underlying sarcoidosis is around 81-98 percent, with an AUC of 0.76 (fair) and 0.96 (excellent). 27
Interestingly, patients with extrapulmonary involvement have been shown to have relatively high levels of serum sIL-2R, suggesting value as a staging and/or severity biomarker. Patients with more advanced radiographic stages and progressive disease also show higher levels of sIL-2R. Serum sIL-2R tests have the highest ability to determine pulmonary severity in comparison to soluble angiotensin-converting enzyme (sACE). Furthermore, in contrast to sACE, an advantage of sIL-2R measurement is that interpretation is not confounded by the use of drugs or immunosuppressants. 27
When using sIL-2R levels as a biomarker for sarcoidosis, it should be noted that an increase in serum sIL-2R values is not always disease specific; elevated values can also be found in other conditions, including hematologic malignancies, other granulomatous diseases, various autoimmune disorders and posttransplantation. Renal insufficiency may also have a major impact on sIL-2 levels, which could lead to misinterpretation of test results. 27 However, even given these caveats, and especially when considering the broader clinical picture of the individual patient, serum sIL-2R measurement is a very useful
prognostic marker. High serum sIL-2R levels can predict the need for therapy in sarcoidosis patients, and high sIL-2R at initiation of therapy has even shown value as a predictor of relapse after therapy with infliximab. 27 Changes in concentration of serum sIL-2R have also been shown to be related to clinical changes, correlating well with changes in pulmonary function parameters and radiological abnormalities. Finally, the serial measurement of serum sIL-2R during disease follow-up has proven useful for assessing the evolution of disease activity in sarcoidosis. 27 In conclusion, when it comes to establishing a credible tool for diagnosing, monitoring and treating sarcoidosis, sIL-2R measurement is here to stay.
The future of sIL-2R measurement
We have seen that sIL-2R measurement can be an extremely useful tool for clinicians to assess immune function in vivo across a broad spectrum of diseases, and established its key role in the investigation and management of sarcoidosis. The adoption of sIL-2R measurement as standard in the clinical environment will, of course, depend on the development of IVDR-compliant assays. However, much of the groundwork in preparation for their debut in the clinic is complete, and sIL-2R ELISA kits are already available for research. We must now begin to home in on how we can use this ubiquitous and complex biomarker to help in the diagnosis or therapeutic monitoring of autoimmune diseases.
References
1. Liao W, et al. Current Opinion in Immunology, 2011, 23 (5), 598-604.
2. Thi Hong Nguyen C, et al. Journal of Dermatology, 2017, 44 (7), 789-797.
3. Peerlings D, et al. Journal of Translational Autoimmunity, 2021, 100123.
4. Barak V, et al. Journal of Autoimmunity, 2009, 33 (3-4), 178-182.
5. Litzman J, et al. Clinical and Experimental Immunology, 2012, 170 (3), 321-332.
6. Lin M, et al. Annals of Hematology, 2017, 96 (8), 1241-1251.
7. Durda P, et al. Arteriosclerosis, Thrombosis, and Vascular Biology, 2015, 35 (10), 2246-2253.
8. Karim AF, et al. Mediators of Inflammation, 2018, 6103064.
9. Damoiseaux J. Clinical Immunology (Orlando, FL), 2020, 218 , 108515.
10. Dik WA, Heron M. Netherlands Journal of Medicine, 2020, 78 (5), 220-231.
11. Rubin LA, et al. Journal of Immunology, 1985, 135 (5), 3172-3177.
12. Rubin LA, Nelson, DL. Annals of Internal Medicine, 1990, 113 (8), 619-627.
13. Rubin LA, et al. Journal of Immunology, 1986, 137(12), 3841-3844.
14. Pedersen AE, Lauritsen JP. Scandinavian Journal of Immunology, 2009, 70 (1), 40-43.
15. Vanmaris RMM, Rijkers GT. Official Journal of WASOG, 2017, 34 (2), 122-129.
16. Yang ZZ, et al. Blood, 2011, 118 (10), 2809-2820.
17. Maier LM, et al. Journal of Immunology, 2009, 182 (3), 1541-1547.
18. Taniguchi T, Minami Y. Cell, 1993, 73 (1), 5-8.
19. Alende-Castro V, et al. All Life, 2023, 16 (1), 2169958.
20. Halacli B, et al. Journal of Critical Care, 2016, 35 , 185-190.
21. Mariotti S, et al. Clinical Endocrinology, 1992, 37(5), 415-422.
22. Manoussakis MN, et al. Lupus, 1992, 1 (2), 105-109.
23. Schimmelpennink MC, et al. Expert Review of Respiratory Medicine, 2020, 14 (7), 749-756.
24. www.nhs.uk/conditions/sarcoidosis
25. Eurelings LEM, et al. PloS One, 2019, 14 (10), e0223897.
26. Vorselaars AD, et al. Respiratory Medicine, 2015, 109 (2), 279-285.
27. Milou C, et al. Sarcoidosis: A Clinician’s Guide (2019), Chapter 19Biomarkers in Sarcoidosis, 219-238.
Tecan recently teamed up with Nature Medicine to host a symposium on the latest ground-breaking research into cancer diagnosis and treatment, from novel biomarkers to personalized vaccines and cell-based therapies. A select group of industry leaders and key experts in oncology came together for a two-day event in Boston, Massachusetts, to accelerate research and clinical discoveries through better partnering between industry and academia. This summit highlighted Tecan’s dedication to assembling leaders in cancer – along with technologists, vendors and solutions providers – so they can share knowledge and expertise, and work collaboratively to battle cancer.
The event was opened by Achim von Leoprechting, Chief Executive Officer at Tecan, who asserted the company’s ambitions to accelerate cancer discoveries in partnership with those in the room. “This symposium is more than an event,” Achim commented. “It’s a demonstration of our joint commitment to driving innovation in oncology, with the ultimate goal of improving patient outcomes and alleviating the pressures on healthcare systems around the world.”
Much of the discussion focused on the need to detect cancer earlier to improve the likelihood of treatment success. Grail, a healthcare company based in California, provided an industry perspective on the subject.
The company is developing a liquid biopsy approach to multi-cancer early screening and diagnosis, which searches the blood for fragments of cell-free DNA released by tumors, detecting more than 50 malignancies in a single test. Jeffrey Venstrom, Chief Medical Officer at Grail, explained that the industry is ready to listen, learn and prioritize areas where joint efforts could improve solutions for laboratories or clinical practices, moving us closer to the dream of standardized multi-cancer blood screening. He also highlighted the challenges associated with transitioning from research to commercialization, emphasizing the importance of automation to ensure consistent and efficient results as a company scales.
One of the challenges for both academia and industry is to ‘democratize’ cancer research efforts, by gathering data from more diverse ethnic backgrounds. Sasha Gusev, a statistical geneticist at the DanaFarber Cancer Institute (DFCI) in Boston, discussed how his work has found that differences in genetic ancestry can bias the results of common assays used to guide lifesaving immune cancer therapies. Addressing this variable in existing biomarkers would bring more effective treatments to a broader population, as well as help to identify those patients that could gain the most from immunotherapy at an earlier stage. This field will likely play a more central role in the future fight against cancer,
which already includes standard treatments for some malignancies.
Darrell Irvine, from the Koch Institute for Integrative Cancer Research at MIT, covered this topic in detail, highlighting the need to expand on existing immunotherapies that treat cancers of the blood, bone marrow and lymphatic system. His research into nanoparticles and synthetic materials is striving to do exactly this, and will hopefully help myriad patients currently not benefitting from some immunotherapies – such as checkpoint inhibitors – by harnessing the power of the immune system to fend off cancer.
Catherine Wu, who leads the division of Stem Cell Transplantation and Cellular Therapies at DFCI, talked about the exciting advances in personalized cancer vaccines.
Catherine’s team analyzes neoantigens – tumor-specific proteins – to generate vaccines that can train the immune
system to attack cancerous cells, leaving healthy tissues unharmed. This fascinating approach is now ready to scale up, which comes with a variety of challenges – including choosing suitable target proteins, optimizing vaccine delivery methods and extending the concept to a broader range of cancers – but is receiving growing interest from investors and pharmaceutical companies, helping to fund further research.
Alanna Church, a molecular pathologist at Boston Children’s Hospital, expanded on the need for a more personalized approach to cancer care, particularly in pediatric patients. There is still a lack of unique pediatric cancer treatments, which is a significant obstacle for oncologists and can sometimes translate to serious consequences for patients. Alanna’s research focuses on the clinical implications of molecular tumor
profiling, which can help clinicians to choose the best therapeutic pathway for young patients. She highlighted the use of the latest technologies – such as next generation sequencing – to accurately characterize known tumors or identify cancer in patients before symptoms appear.
Every tumor is a ‘malignant snowflake’ with its own molecular profile and biological characteristics according to Vivek Subbiah, an oncologist and former medical director of the Clinical Center for Targeted Therapy at MD Anderson Cancer Center, who now works as the Chief of Early-Phase Drug Development at the Sarah Cannon Research Institute in Nashville, Tennessee. He illustrated that even rare cancer types can now be treated with the aid of tumor genome sequencing and targeted drugs, using RET gene mutations and fusions as examples.
These industry leaders, clinicians, academics and researchers are shaping the future of oncology. However, their brilliance alone cannot drive the discoveries to dramatically improve patient outcomes, and collaboration is the essential element to progress cancer screening, diagnostics and treatments. Tecan will once again host industry experts, pioneering researchers and leading oncologists in 2024, to create strong partnerships that push the boundaries of research and accelerate innovation in oncology.
Large structural variations in the genome are responsible for many diseases and conditions, including cancers and developmental disorders. Gene changes – including insertions or deletions, translocations, inversions and duplications – can lead to alterations in how and when a gene is expressed, impacting on a wide range of in vivo processes. Bionano Genomics has developed an optical genome mapping platform offering high speed, high throughput whole genome mapping to support genomic research into human disease.
Bionano was established as a spin-out from Princeton University in 2003, and now employs over 300 people across sites in the US, Canada and Europe.
Dr Erik Holmlin, president and chief executive officer, explained: “Bionano is a genomics company focused on human health and wellness. We recognize that the application of genomics is primarily focused on driving healthcare forward, from the discovery of new drug targets to
improving diagnostics and developing precision medicine approaches. Broader than that, we also see the value of genomics in day-to-day wellness, giving people access to their genomic data to help them make better health decisions and take preventive actions.”
Bionano’s technology platform, the Saphyr® system, uses optical mapping to detect structural variations in the genome. Erik added: “Next generation sequencing cannot answer all of the questions that clinicians and pathologists are seeking to address. We focus on the structure of the genome, as it is far more sophisticated than just some letters lined up like the alphabet. It really matters how things are organized in the genome: what chromosome the gene is on; how many copies of the gene are present; what kind of signaling information is ahead of the gene; and what results in it being turned on in some cells and not in others. That’s a problem that’s very difficult to solve with existing tools, and so we developed a methodology, instrumentation and the reagent kits needed to view the structure of the genome and discover the impact of structural changes on numerous healthcare questions.”
“When we started to develop the Saphyr system in late 2014, we already had the IRYS system, which was our first generation optical genome mapper,” Erik continued. “It was a powerful system, but had many limitations, including its speed and cost, as well as its overall performance in terms of mean time between failures. However, it was a great learning
platform for us to see what we needed to develop in a new instrument. It had to be faster, more easily manufacturable, cheaper for the end user, and more reliable.”
“We had the capability to develop the system in house, but needed a partner with experience in scalability and a global supply chain to support manufacturing. We were looking for a company that had experience in complicated instrumentation, as well as knowledge of medical and clinical products, because of the complex regulatory landscape in those areas. One of the things that made Paramit stand out from other providers was the sophisticated medical systems that the company was building at the time. Our COO had also worked successfully with Paramit on previous projects, and that level of confidence and trust is so important on a big project.”
“We undertook a majority of the design ourselves, then worked closely with Paramit to bring the instrument to life. We relied heavily on the Paramit team’s materials knowledge and supply chain awareness to ensure the system we were developing could be built predictably, in the required quantities and timelines. The aim was to create a more reliable and affordable machine that we could deliver dependably and,
with the support of Paramit, we’ve consistently hit our objectives. The first generation of the Saphyr system launched in 2017 and, since then, we have made a number of incremental improvements over time, leading to the second generation Saphyr system that is currently on the market.”
Erik concluded: “Paramit has been there to support us throughout the development process, and we’ve shared a win-win mentality with the team there. They’ve taken the time to understand our design goals and vision, then we’ve worked together to turn those into the successful product available today. It’s really been a good partnership based on a collaborative approach between multidisciplinary teams striving to meet common end goals.”
For research use only. Not for use in clinical diagnostics.
To find out more about Paramit, visit www.paramit.com
To learn more about Bionano, go to www.bionanogenomics.com
The aim was to create a more reliable and affordable machine that we could deliver dependably and, with the support of Paramit, we’ve consistently hit all of our objectives.The Saphyr system is a genome imaging tool for high throughput structural variant detection and analysis
NGS is a vital tool used for studying the structure and function of DNA for multiple applications. However, there are several challenges commonly encountered when using this technique, particularly when working with degraded or trace levels of RNA. These issues motivated research staff at Kazusa DNA Research Institute in Japan to search for library preparation kits that would enable high quality sequencing for its customers when working with low quality samples.
Kazusa DNA Research Institute (KDRI) in Kisarazu, Chiba Prefecture, was established in 1994, making it the world’s first organization specializing in DNA-related research. Since then, the institute has been involved in many human and plant genome projects, and is recognized as a global leader in genetic studies. It performs NGS on human, plant and bacterial samples provided by clients from diverse
sectors. This includes the agriculture industry – where genetic analysis is crucial for breeding commercially important plant species with desired traits, and for standardizing the quality of plant seeds – and medicine, where KDRI runs NGS to support the early detection of rare diseases and the identification of novel cancer markers and precursors.
KDRI assessed the capability of the Revelo™ RNA-Seq High Sensitivity library preparation kit to process low-input and degraded or trace level RNA samples prior to NGS. This flexible, end-to-end library preparation solution produces rRNA libraries from 10 250 pg to 10 ng of total RNA using Tecan’s proprietary SPIABoost™ technology. Dr Kazuto Kugou, a researcher in the Laboratory of Gene
Sequencing Analysis at KDRI, explained why the center made this decision: “We chose to trial the Revelo kit because it was specifically formulated for the preparation of highly degraded samples – for instance, samples with an RNA Integrity Number of just 1.8 – and those containing only trace amounts of RNA. We frequently encounter these types of specimens, and typically handle low throughput applications, so the Revelo system seemed like the perfect solution for us and we were keen to try it out.”
Dr Kugou described some of the advantages of the Revelo kit over the alternative options they had previously tested in the lab: “The library preparation tools offered by other companies all require an additional DNA purification step in order to remove adapter dimers. In contrast, the Revelo solution includes DNase treatment, and this one simple step is enough to remove all the adapter dimers completely. The kit therefore enabled us to prepare a large number of clean libraries within a short timeframe using only a few PCR cycles, reducing PCR bias and providing unbiased 5’ to 3’ transcript coverage.”
Part of the lab’s role is to sequence RNA-Seq libraries and map the sequencing data onto a human reference sequence. This had proven
difficult in the past when using poor quality specimens, but the team noticed that sufficient quality data for analysis was obtained during the trial of the Revelo kit. “We are always concerned about the mapping rates we will achieve from samples with highly degraded or trace level RNA, but when we used the Revelo system to prepare the libraries, we observed unique mapping rates of over 80 percent,” said Dr Kugou.
“Our institute aims to be a world leader in all aspects of DNA research, and we strive to contribute to society through medicine, agriculture, industry and education. We are excited to see what new breakthrough solutions the company develops over the coming years to further advance the field of genetic analysis,” Dr Kugou concluded.
For research use only. Not for use in clinical diagnostics.
To find out more about the Revelo RNA-Seq High Sensitivity library preparation kit, go to lifesciences.tecan.com/revelorna-seq-high-sensitivity
For more information about the Kazusa DNA Research Institute, visit www.kazusa.or.jp/en
We chose to trial the Revelo kit because it was specifically formulated for the preparation of highly degraded samples… and those containing only trace amounts of RNA… the Revelo system seemed like the perfect solution for us...
High throughput imaging of cells is a key part of immunological research, allowing the visualization of the mechanisms of action and in vivo effects of infectious viruses to aid the development of effective antiviral treatments and vaccines. Chinese biotechnology company FantasiaBio is using automated multicolor imaging to accelerate its development of innovative technologies for immunodiagnostics and vaccine development.
FantasiaBio, based in the Jinhua region of China’s Zhejiang province, understands the power of imaging live cells, and has created a range of fluorescence-based tools to exploit the power of multiplexed live cell imaging for clinical research and development. In 2020, the company developed an innovative in vitro assay to quantify
neutralizing antibodies against SARSCoV-2, allowing researchers to evaluate an individual’s level of protection after administration of a COVID-19 vaccine.
Qin Xiao-Feng, Co-founder and Chief Scientific Officer at FantasiaBio, explained: “The kit uses a vesicular stomatitis virus (VSV) as a pseudotype vector with a green fluorescent protein
(GFP) payload. If the COVID-19 vaccine stimulates effective production of neutralizing antibodies, the VSV pseudovirus is unable to infect human target cells, so the GFP gene is not expressed in these cells. The expression of GFP can normally be seen using a fluorescence microscope, and the number of green ‘dots’ is inversely
proportional to the antibody activity. However, imaging in this way requires researchers to be sitting in front of a microscope for long periods of time to manually assess samples, which is time consuming, laborious and leads to inter-operator variability in results. We therefore needed a way to streamline and accelerate whole-well imaging for multiwell plates.”
Qin continued: “We are a technologydriven company, and routinely perform two-, three- or even four-color multiplex studies for up to 100 microplates a day, so automation is essential. Automated microscopes have come a long way over the last few years, but these high resolution systems have limited throughput, so we needed a better balance between speed and resolution. When Tecan introduced the Spark® Cyto in the China market, we immediately recognized that it was exactly what we were looking for. We did a quick proof-ofconcept study to perform high throughput testing of our neutralizing antibodies assay, and found it to be a good fit for our needs, so we purchased a system in October 2020.”
“The main feature of the Spark Cyto that was attractive to us was its combination of wide field imaging and high imaging speed. We use the low magnification lens – 2x or 4x – which we have found to be sufficient to obtain
quality images and accurate single cell counts. Thanks to the Spark Cyto, we now have the capacity to rapidly image an entire 96-well plate, with just one single frame to cover the entirety of each well, avoiding the need to stitch together multiple frames for analysis.”
“The system is also proving invaluable for screening and evaluating COVID-19 vaccine candidates. Our aim is to develop a new generation of vaccines that prompt a stronger and longerlasting immune response – mucosal as well as T cell activation – that will lead to the prevention of infection, instead of helping to reduce the symptoms or mortality. This means that we do a lot of routine work, such as viral titer assays and precisely measuring antibody titers (ID 50) in samples collected from various anatomical locations, which is well suited to an automated instrument such as the Spark Cyto.”
“The Tecan team has been very helpful, from when we started looking at new imaging instruments to being quick in providing technical support and addressing our specific challenges since purchasing the Spark Cyto. Our local engineer is very helpful, and quickly helps us resolve most issues by phone, or sometimes they just come from Shanghai to rescue us! This local support is very important, and we’re very glad to have it.”
For research use only. Not for use in diagnostic procedures.
To find out more about Tecan’s Spark Cyto, go to www.tecan.com/sparkcyto
To learn more about FantasiaBio, visit www.fantasiabio.com
The Tecan team has been very helpful, from when we started looking at new imaging instruments to being quick in providing technical support and addressing our specific challenges since purchasing the Spark Cyto.
Transitioning from the world’s current petroleum-based industry to a sustainable bioeconomy depends upon the microbial upcycling of plant-based feedstocks for monomer production. Researchers at Forschungszentrum Jülich (FZJ) in Germany have introduced automation platforms to enable the rapid engineering of microbial strains that can convert renewable raw materials into value-added compounds.
FZJ is one of the largest interdisciplinary research centers in Europe, with over 6,000 employees focused on solving major global health and environmental challenges. Dr-Ing Stephan Noack and his Quantitative Microbial Phenotyping (QMP) group in the FZJ’s Institute of Bio- and Geosciences are using high throughput experiments, quantitative omics technologies and process modeling to optimize the bioproduction of value-added biochemicals, such as organic acids for biopolymers, amino acids as feed and
food additives, and high value pharma ingredients that can be manufactured on a near-industrial scale. The team’s workflows typically start with incorporating different genetic ‘pieces’ into plasmids via standard cloning techniques, for instance Golden Gate or Gibson assembly. This genetic material is then transformed into model organisms like Escherichia coli, Corynebacterium glutamicum or Pseudomonas putida, which are subsequently plated onto agar plates. Colonies are later picked and prepared
for characterization in liquid culture. Finally, strain performance is analyzed in detail using small-scale cultivation and quantitative mass spectrometry.
Automation plays a central role in these workflows, and Stephan explained the reasoning behind the lab’s decision to automate: “Creating and testing novel producer strains with complex biosynthetic pathways is extremely slow and error prone when performed manually. We therefore wanted to introduce automation for all of our molecular cloning and strain characterization steps in order to streamline our work, increase lab productivity, optimize assay precision and consistency, and relieve operators from tedious repetitive tasks. We started our automation journey in 2008, when we approached Tecan to set up an automation platform for the rapid phenotyping of existing strain libraries. This ‘mini pilot plant’ can automate individual phenotyping steps – such as media preparation and optimization, inoculation and timeor signal-triggered sampling – and includes a microbioreactor device to enable well-controlled cultivation in microplates. Tecan was – and still is –the only laboratory automation supplier that could guarantee the continuous shaking of microbioreactor cultures without stopping for sampling, so it was an obvious choice to partner with the company.”
The QMP group took another step towards complete automation of the design-build-test-learn cycle by acquiring a Fluent® 1080 Automation Workstation at the beginning of 2021, enabling faster construction of microbial strains and a shorter turnaround time to viable bioprocesses. This system features an integrated Resolvex® M10 positive pressure solid phase extraction system for preparing samples, and a Pickolo™ Colony Picker for hands-free clone selection. It also includes a centrifuge, an orbital shaker, a cooling carrier and a barcode scanner.
Stephan’s colleague in the QMP team, Dr Julia Tenhaef, described how the new system fits into their lab: “Right from the beginning, we had an open discussion with the Tecan team regarding our exact requirements, and they designed a custom solution that fulfils all the needs of our sophisticated workflow, meaning that strain construction is now almost entirely automated. The installation and set-up was quick and straightforward, allowing us to transition seamlessly onto our new platforms and minimize downtime. As you’d expect with such a highly integrated system, we have had some issues, but Tecan’s application
specialists are always on hand to provide help and feedback whenever we have questions.”
“The Fluent is truly multi-functional, enabling us to carry out a wide variety of tasks on just one instrument, which is really convenient and efficient,” Julia added. “For example, we can perform fast and precise liquid handling, easily switching between different labware formats and workflow steps with little or no operator interaction. It’s also compact and integrates easily with the other instruments that we already had in the lab, which is a huge plus.”
She continued: “We use the NucleoSpin 96 Plasmid Core Kit from Macherey-Nagel in combination with the Resolvex system for plasmid isolation. After cell lysis, the cell debris and chromosomal DNA is removed from the samples via filtration, then the plasmid DNA is bound to the Macherey-Nagel plasmid binding plate. Subsequent washing steps are also performed using the Resolvex instrument, and the final elution of the purified plasmid is done using the integrated centrifuge. We received valuable support from MachereyNagel when establishing the ideal pressure and time settings needed for
using the kits with our set-up, and Tecan helped with creating and optimizing custom movement vectors for the Fluent to guarantee flawless loading and unloading of plates and custom labware.”
“We’re extremely satisfied with the Fluent and Resolvex integration, and we haven’t even come close to the maximum throughput of the workflow yet, so we definitely have a lot of room to grow and increase our productivity. We’re excited to take advantage of the benefits our Tecan automation solutions have to offer for this type of research, and we’re confident they will be crucial to our success in the years ahead,” concluded Stephan.
To find out more about Tecan’s cell biology solutions, go to lifesciences.tecan.com/ applications_and_solutions/ cell_biology
For more information on Forschungszentrum Jülich, visit www.fz-juelich.de/en
The Fluent is truly multi-functional, enabling us to carry out a wide variety of tasks on just one instrument…
The prevalence of eye disease is rising around the world and, for most of them, there are no effective therapies available. Disorders that impair vision – such as macular degeneration or glaucoma – are a leading cause of disability and loss of an independent lifestyle in aging populations. At the other end of the spectrum, the incidence of myopia – or short-sightedness – is also on a steep incline, with up to 90 percent of teenagers being affected in some regions. Researchers in Basel are using various cuttingedge tools – including single-cell genomics – to understand the molecular mechanisms behind some of these diseases, with the aim of developing effective therapeutics.
The Institute of Molecular and Clinical Ophthalmology Basel (IOB) was established in 2018 to enable researchers and clinicians to advance our understanding of vision and its diseases, and to develop new therapies for vision loss. As part of its growth, IOB has set up a state-of-theart single-cell genomics facility to provide guidance, support and advice to other groups at the institute wishing to use these techniques in their research. One of the aims of the department is to develop new singlecell methods to improve data quality, increase throughput and reduce costs. Simone Picelli, Platform Leader, explained: “I have been working with single-cell RNA sequencing for many years, trying to come up with new ideas or new protocols that can be automated and miniaturized to reduce the costs and make it more efficient. We have recently developed a novel workflow – FLASH-seq1,2 – that we have also automated with Tecan’s Fluent® Automation Workstation.”
Single-cell RNA sequencing has transformed genomics in the last decade, with plate-based methods –
such as Smart-seq2 3 – routinely being used as an alternative to water-in-oil emulsion methods. These approaches offer superior sensitivity, and the ability to provide full-length transcript information, but they typically require extensive hands-on time, leading to lower throughput and a higher cost per sample. Simone continued: “We developed FLASH-seq to address most of these issues, generating sequencingready libraries in a single workday while providing superior data quality.1 We have created an automated version of the FLASH-seq workflow, where throughput is only dependent on reaction and incubation times, manual intervention is limited to the preparation of master mixes, and almost all the dispensing and clean-up steps are performed by the instruments. We have found that automating our single-cell RNA library prep has made big improvements to our throughput, data quality and reproducibility, while also reducing reagent costs and saving time.”
The IOB turned to Tecan when developing the automated workflow, to ensure the liquid handling system
would meet its specific needs. “I have used Tecan equipment for many years,” Simone added. “I started out using a Freedom EVO®, which I really liked for its intuitive software and good amount of deck space. It was also very robust, even in my inexperienced hands at the time. When I moved to Basel, I switched to the newer Fluent Automation Workstation because, while I would have happily kept using the Freedom EVO, I could see clear advantages of moving on to the latest platform.”
Almost every step of the FLASH-seq workflow has been automated –including single-cell isolation, cDNA preparation, purification, quality control and quantification, and NGS library preparation – to simplify plate handling. “We run the protocol using 384-well plates, and many of the steps are automated using the Fluent. There are several stages where liquid needs transferring or adding to every well, and this can be achieved in a single step with the 384-channel head on the Multiple Channel Arm™, so it only takes five seconds for a whole plate. This arm can pipette down to 300 nanoliter volumes for aqueous solutions, which is
ideal for transferring DNA or primers. We also need to add index adaptors to the different cells, which we can do easily with the Fluent. There is enough space on the deck to store plates and tip boxes, which is very convenient and it allows us to process multiple plates in parallel. We also have an integrated shaker for steps which require the use of magnetic beads, as these need to be continuously agitated to stop them from settling.”
Automating single-cell RNA sequencing workflows allows researchers to minimize hands-on time, improve reproducibility and manage costs, and miniaturization can take these savings even further. Simone concluded: “I have worked with Tecan software for many years, and am comfortable developing more and more complicated scripts, as I know that the system is very reproducible and robust. This has allowed us to develop a very efficient automated FLASH-seq workflow, detecting many more genes, much more easily, compared to other published methods.”
We have found that automating our single-cell RNA library prep has made big improvements to our throughput, data quality and reproducibility…
1. Hahaut V. et al. Fast and highly sensitive full-length single-cell RNA sequencing using FLASH-seq. Nat Biotechnol, 2022, 40, 1447-1451. doi:10.1038/s41587-02201312-3
2. Picelli S. and Hahaut V. FLASH.seq protocol V.4. protocols.io, 2023. doi:10.17504/protocols.io.kxygxzkrwv8j/v4
3. Picelli S. et al. Smart-seq2 for sensitive full-length transcriptome profiling in single cells. Nat Methods, 2013, 10, 1096-1098. doi:10.1038/nmeth.2639
To find out more about Tecan’s automated NGS library preparation solutions, visit lifesciences.tecan.com/ngssample-preparation
For more information about the IOB single-cell genomics platform, visit www.iob.ch/research/molecularresearch-center/single-cellplatform-s-picelli
Today’s drug discovery and development pipelines rely heavily on biosensors for screening and characterizing candidates, and many of these workflows are based on surface plasmon resonance (SPR) technology. SPR is an optical technique that uses polarized light to generate electron waves (plasmons) on an electrically conducting surface at the interface between two media. Creation of plasmons reduces the intensity of reflected light at a specific angle – known as the resonance angle – which depends directly on the refractive index of the sensor surface. Molecules binding to this surface will change its refractive index, making it possible to monitor molecular interactions without labeling the reactants. One binding partner can be immobilized to the surface, while the second binding partner is injected across the sensor. When complexes
are formed, the change can be registered by the SPR sensor, providing information about binding events, including the association and dissociation reaction kinetics.
Conventional SPR platforms commonly use unidirectional flow, which limits association and dissociation times, imposing restrictions on kinetic studies and increasing the sample volumes required for screening. Tim Germann, Chief Commercial Officer at Carterra, commented: “Here at Carterra, we have changed the analytical paradigm by developing a proprietary solution that uses a network of microchannels, enabling bidirectional flow-based array printing of molecules on sensor surfaces. These microchannels flow and cycle ligand solutions over a surface, reducing the sample volumes needed and dramatically increasing
throughput by addressing 384 ligands in a single run. Analyzing so many samples in parallel accelerates screening 100-fold compared to legacy technologies, which is invaluable for the pharmaceutical industry.”
Carterra partnered with Paramit – a Tecan Group company – to manufacture the LSA platforms. Tim explained: “We chose Paramit as our contract manufacturer to produce our instruments because of its strong reputation and vPoke® Mechanical Assembly – a highly accurate system that allows fine control of manufacturing parameters. We first approached Paramit in 2017, and were impressed by its production facility, as well as the vPoke system itself. We worked closely together on the technology transfer to manufacturing, and had production units going out to
customers by the end of 2018. Based on feedback from these early customers, we were able to improve the overall product offering and reliability. We have nothing but positive things to say about this collaboration.”
“Our LSA platform has already been part of a great success,” Tim continued. “Its exceptional speed and efficiency enabled the development of the world’s first COVID-19 therapeutic –bamlanivimab – by Eli Lilly in just 90 days. This highly efficient screening method is also very useful for AI-driven drug discovery pipelines, which can generate as many as 5,000 antibodies that need to be screened and characterized in a week. Companies involved in this type of modern
therapeutic research can use our system every day to rapidly characterize their candidates, and use the resulting data to inform and refine their models.”
“Since launching the LSA platform, we have attracted customers across the globe – from tiny start-ups to large, well-established pharmaceutical companies – that have all seen the value in getting more information faster and earlier in the discovery process. Aside from the remarkable increase in throughput, our technology provides a greater level of detail regarding binding interactions, including information on
affinity and epitopes. Just as the Hubble Space Telescope enabled the discovery of millions of new galaxies, users of Carterra’s platforms can see things at a time and resolution that was not previously possible. That’s the definition of transformative,” Tim concluded.
For research use only. Not for use in clinical diagnostics.
To learn more about Tecan’s vPoke Mechanical Assembly technology, go to www.paramit.com/vpokemechanical-assembly
For more information about Carterra, visit www.carterra-bio.com
Just as the Hubble Space Telescope enabled the discovery of millions of new galaxies, users of Carterra’s platforms can see things at a time and resolution that was not previously possible.
Diagnosing hormone-based disorders using saliva samples dates back to the 1980s, but it is only the sensitivity improvements over the last decade that have led to saliva-based diagnostics becoming a viable alternative to blood testing. Biovis’ Diagnostik, a medical laboratory based in Limburg, Germany, has been at the forefront of saliva diagnostics since 2012, providing comprehensive testing to improve the analysis of hormone-based disorders.
Biovis’ Diagnostik was founded in 2004 to offer a broad range of clinical diagnostic tests, including gastrointestinal, immune and metabolic analyses. The company began looking into the benefits of saliva testing in 2010, after requests from many smaller labs for support. Dr. med. Burkhard Schütz, founder of Biovis’, explained: “I had always worked with serum diagnostics, and there were always a few reasons that spoke against saliva diagnostics to me. However, when I started to look into it, I realized that it was actually extremely interesting, and could provide complementary findings to support more traditional diagnostics. We therefore started to offer more services in this area, putting us on the journey to becoming one of the largest providers of saliva hormone testing in Germany.”
“Over the last 10 years, saliva diagnostics has increased in popularity as an alternative to blood-based testing, and has now established itself as a stable field – I would have made a big mistake if I hadn’t taken it up,” Burkhard continued. “It offers an incredibly simple way of collecting samples. It’s non-invasive, and the patients can take samples themselves, without the need to go to the doctor’s office. They can take the samples at any time across the day, which is very useful for determining various phase-dependent parameters – such as melatonin or cortisol – that you can only monitor in blood if the patient comes to the practice again and again. The stability is also very good, and samples are easier to transport than blood.”
Saliva diagnostics is perfectly suited to the analysis of biologically active, free hormone levels, offering accurate diagnosis of hormone disorders –particularly where regular testing is required. Hormone levels can be monitored using simple immunoassays, which can be easily automated when they are being carried out on a large scale. “These benefits make it highly advantageous to measure various types of hormones in saliva, including cortisol, melatonin, progesterone, estradiol and testosterone. This approach is particularly useful in several phasedependent disorders, such as monitoring progesterone and estradiol throughout the menstrual cycle, to be able to correct imbalances associated with pre-menstrual syndrome. It is also very helpful for detecting adrenal gland weakness, by tracking changes in cortisol and other adrenal hormones. A third use is for monitoring estriol levels, since imbalances can be associated with dry mucous membranes and frequent urinary tract infections. All of these imbalances can be corrected with therapies once they have been identified correctly, so accurate diagnosis is crucial.”
Biovis’ has streamlined its saliva testing activities by integrating Tecan hormone diagnostics ELISA kits into its workflow. The lab uses three Freedom EVOlyzer® platforms to automate all common ELISA steps, including sample distribution and predilution, reagent pipetting, incubation, plate washing, optical density measurements and result generation. These instruments support the running of Tecan ELISA kits for estradiol, estriol, progesterone, cortisol, testosterone, DHEA and melatonin measurement in salivary samples, as well as the quantification of free vitamin D, neopterin and serotonin in serum or urine samples. Automating the process has allowed Biovis’ to increase throughput, while improving the accuracy and reproducibility of results. “I have worked with Tecan for many years, which has built a relationship of trust. The Tecan ELISA kits provide reliable results, and the support is there if any issues occur. Saliva diagnostics is a simple tool, and I believe it will continue to be an important approach for providing better diagnosis of hormone imbalances in cycle disturbances –prolonged or shortened cycles – or the
I have worked with Tecan for many years, which has built a relationship of trust. The Tecan ELISA kits provide reliable results, and the support is there if any issues occur.
absence of ovulation. It can also support the many women who have pre-menstrual syndrome associated with changes in the progesterone-estradiol ratio. Being able to detect these imbalances allows women to seek therapies to overcome their challenging symptoms,” Burkhard concluded.
For research use only. Not for use in clinical diagnostics
For more about Tecan’s saliva diagnostic solutions, visit www.tecan.com/saliva-generalinformation
To learn more about Biovis’ Diagnostik, go to www.biovis.eu/en/
The European Union’s In Vitro Diagnostics Regulation (IVDR) replaces the In Vitro Diagnostic Medical Devices Directive (IVDD), completely overhauling the regulations regarding pre- and postmarket requirements for IVD devices. This has implications for the entire supply chain, from manufacturers with responsibilities for design, development and commercialization, to agreements between manufacturers and key economic operators, such as importers and distributors. The regulation also introduces specific requirements and limitations on hospitals and labs developing their own diagnostic tests, which must review and, if necessary, replace in-house assays with commercially available CE-marked, IVDRcertified alternatives to ensure compliance for the chosen application.
The IVDD came into effect more than two decades ago, and a lot has changed since then. There was a clear need for reform to cover the gaps created by new technologies – such as point-of-care and genomics devices – and products targeting personalized medicine, which are not covered by the IVDD. This led the European Union to embark on a major regulatory overhaul to modernize device classification, and address any product safety or quality issues that may have gone unnoticed. The resulting IVDR came into force in May 2017, requiring all existing IVD devices, plus any new devices coming to market, to meet the requirements of the new legislation. The transition period was initially due to end on May 26, 2022 – a tight deadline that was compounded by the COVID-19 pandemic – and so a more progressive roll out was implemented to prevent disruption in the supply of essential healthcare products. While all new IVD devices and Class A non-sterile devices were required to be IVDR compliant from May 2022, a more gradual implementation –between 2025 and 2028 – is now permitted for devices in other classes and in-house assays.
The IVDR contains very specific requirements for analytical and clinical performance, and every manufacturer must demonstrate that its devices meet these standards, as well as the general safety performance requirements for device safety and quality. There is also an increased need for post-market vigilance activities to ensure cradle-to-grave compliance, with clearly defined responsibilities for
manufacturers, distributors, importers and authorized representatives to enhance transparency throughout the entire supply chain. The benefits are evident, but what is less obvious is the effect on commercial labs creating their own in-house tests, who are now defined as manufacturers and must therefore comply with IVDR. While it may, in some cases, be possible to switch to a CE-marked test, commercial assay manufacturers are likely to be reluctant to invest significantly in niche products with a limited market.
Ganzimmun Diagnostics in Mainz is one of the largest labs in Germany, serving both primary care doctors and complementary medicine practitioners. It offers a wide variety of genetic, clinical chemistry and immunoassay tests for different clinical indications, some of which have been developed in house. Ganzimmun’s PRRC (person responsible for regulatory compliance) Petra Hammer explained the impact of IVDR on her laboratory: “Under IVDR, we are both a customer using thirdparty assays and a manufacturer of lab-developed tests. We use a range of commercially available assays from Tecan – predominantly saliva diagnostics, but also other endocrinology and immunology assays, as well as complementary tests for integrative medicine – which we run on a Freedom EVO® platform, giving us a complete solution for high throughput analysis. In addition, we use a large number of in-house tests, all of which must be reviewed, and any necessary action taken to comply with IVDR, by the end of 2028.”
Petra continued: “We began by performing a gap analysis to establish the difference between IVDD and IVDR, to see what we needed to do to ensure our existing assays were compliant and could continue to be used. Most of the tests we use are Class B, with a few in Class C. We can’t discontinue in-house tests that are still required by our clients when there is no CE-marked alternative available, so we have to check whether the validation of these assays is acceptable for IVDR. We found that around 20 percent of our assays –generally Class C tests – need revalidating, which will require a lot of time and resources. The biggest challenge for us is that IVDR demands technical documentation of in-house software. As this was not necessary before, and much of the work done in the past is not documented, so we have to establish what was done, when, and how. We are also obliged to perform post-market surveillance now, which consumes further resources.”
Every department at Ganzimmun offers a different range of in-house assays, and it is each team manager’s responsibility, with support from the quality management team, to establish
which tests – if any – must be revalidated. “In the past, tests were developed in each lab, with no specific department to guide the process. To overcome this, we recently set up a department with responsibility for assay development. This team supports the individual labs by providing information to help them devise a plan for revalidation, testing and statistical analysis.”
While Ganzimmun is responsible for ensuring its in-house assays are IVDR compliant, for third-party tests the burden falls on the manufacturers, who must work with a Notified Body to achieve IVDR certification for their products. As a result, new tests required to extend the scope of a lab’s services may not be available as soon as they would like; it can be quicker to develop an assay in house, as this does not require a Notified Body. There is much to be gained by clinical laboratories and IVDR-ready manufacturers – such as Tecan –collaborating to ensure that labs have access to CE-marked tests that meet their needs. “Labs need tests that are both suitable for their applications and regulatory compliant. CE-marked tests validated by the manufacturer are
IVDR compliant by definition, but we still need to check that they work as expected with our samples and the equipment in our lab. If we are uncertain whether the way we are using a test complies with the manufacturer’s intended purpose, we ask them to help us check that we are performing the assay correctly. The continuous communication between our lab and Tecan gives us the information and practical support that we need, as well as allowing us to provide input about our needs and challenges, with benefits for both parties,” Petra concluded.
For more about Tecan’s IVDRready assays, visit www.tecan.com/ivdr-overview
To learn more about Ganzimmun Diagnostics, go to www.ganzimmun.de
We use a range of commercially available assays from Tecan… which we run on a Freedom EVO platform, giving us a complete solution for high throughput analysis.
Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276
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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
