by Professor Paul Kaye Leishmaniasis is classified as a neglected tropical disease. It is the cause of a huge health burden and is common in Asia, Africa, South and Central America, and even southern Europe. This article discusses how flow cytometry can help to evaluate diagnosis, monitor the effects of therapy and help in the creation of a vaccine.
Background
The leishmaniases are a family of devastating diseases, affecting a great many people across the globe and presenting a significant risk to both public health and socioeconomic development. The leishmaniases are vector-borne diseases, caused by infection with one of 20 species of the parasitic protozoan Leishmania (Fig. 1), transmitted through the bite of the infected female phlebotomine sand fly.
They can be broadly classified as tegumentary leishmaniases (TLs), affecting the skin and mucosa, and visceral leishmaniasis (VL), affecting internal organs. Whereas VL is responsible for over 20¦000 deaths per year, TL are non-life-threatening, chronic and potentially disfiguring, and account for around two-thirds of the global disease burden.
Within TL, there are three subtypes: self-healing lesions at the location of sand fly bite (cutaneous leishmaniasis; CL), lesions that spread from the original skin lesion to the mucosae (mucosal leishmaniasis; ML), and those which spread uncontrolled across the body (disseminated or diffuse cutaneous leishmaniasis; DCL). VL, also known as kala azar, involves major organs including the spleen, liver and bone marrow. In addition, patients recovering from VL after drug treatment often develop post kala-azar dermal leishmaniasis (PKDL), a chronic skin condition, characterized by nodular or macular lesions beginning on the face and spreading to the trunk and arms. As it may develop in up to half of patients previously treated and apparently cured from VL, it is thought that PKDL plays a central role in community transmission of VL.
The World Health Organization designates leishmaniasis as a neglected tropical disease (NTD), which together affect more than one|billion people across 149 countries worldwide; true prevalence may be even higher. Disproportionately, NTDs affect the poorest, malnourished individuals, and contribute to a vicious circle of poverty and disease. The significant physical marks, including ulcers, often left in the wake of the TLs may have an impact on mental health and perpetuate social stigma associated with the diseases [5]. There are over 1|million new cases of TL and 0.5|million new cases of VL each year, which together account for the loss of approximately 2.4|million disability-adjusted life years.
Treatment challenges
Leishmaniasis treatment can be quite difficult since at-risk populations may lack access to healthcare, and the limited battery of drugs has been increasingly compromised by resistance. Additionally, because the parasites in question are eukaryotic, they are not dissimilar from human cells, so the medication is also liable to be harmful – even fatal – to host as well as to pathogen.
Although the burden of VL in South Asia has been reduced with single-dose liposomal amphotericin B, the drug is less effective in other geographic locations, namely East Africa. Various drug combinations have been tested, unsuccessfully, and new chemical entities and immune-modulators are in early stages of development and as yet untested in the field. Unfortunately, little has changed in the treatment for CL for the past 50|years.
No vaccines are currently approved for any form of human leishmaniasis, although vaccines for canine VL have reached the market. Barriers to vaccine development include the limited investment in leishmaniases R&D and the high costs involved in bringing new products to those that need them.
Current work
My work on leishmaniasis has taken a holistic view, rooted in the immunology of the host-parasite interaction, but employing tools and approaches that span many disciplines: mathematics, ecology, vector biology and most recently neuroscience. Thirty years of discovery science has led to the development of a candidate for a therapeutic vaccine for PKDL, the mysterious sequela to VL [6]. ‘Therapeutic’ vaccines are given after an individual is infected with a pathogen and are designed to enhance our immune system and help eliminate the infection.
With colleagues from Sudan, we are in the midst of a phase IIb clinical trial funded by the Wellcome Trust, evaluating the efficacy of this therapeutic vaccine in Sudanese patients with persistent PKDL.
However, the research has been a long time in the making and has a long way to go. To continue to make progress, we linked with colleagues in Ethiopia, Kenya and Uganda and at the European Vaccine Initiative (http://www.euvaccine.eu/) in Germany, to develop a new research consortium to evaluate the immune status of people suffering from leishmaniasis. For example, using flow cytometry for blood and multiplexed immunohistochemistry for tissue biopsies, we can enumerate the proportions of lymphocytes, monocytes and neutrophils based on surface marker expression (e.g. CD3, CD19, CD14, CD16), and characterize their function, for instance by expression of cytokines (e.g. interferon-gamma) or other cell surface proteins that define function state. To support this endeavour, we recently received a grant from the European & Developing Countries Clinical Trials Partnership (EDCTP) that will allow us to not only extend our vaccine programme in Sudan [9] but also to address other important research challenges.
To develop vaccines and indeed new drugs, we often need tools capable of performing in-depth comparisons of how the body’s immune system is coping with the infection when a patient is first admitted to hospital and how it changes as the patient undergoes treatment and is hopefully cured. For example, recent evidence suggests that during infection, T lymphocytes may become ‘exhausted’ and unable to fight infection and the exhausted state can be identified by expression of surface molecules such as programmed cell death protein|1 (PD-1) and lymphocyte activation gene 3 protein (LAG-3). It is important to know if exhaustion can be reversed following treatment or whether we need to stimulate new populations of T lymphocytes. By understanding these nuanced changes in immune cells in our blood, we can design ways to improve how vaccines and drugs work in concert with immune cells, and understand why some patients might relapse from their disease or develop PKDL. Flow cytometry is a central tool for immunologists and plays a critical role in uncovering mechanisms of immunity and in assessing how well vaccines work and biomarkers of drug response. It uses antibodies that recognize specific molecules or markers on the surface or inside immune cells, such as those mentioned above, that help us predict their function. These antibodies are fluorescently labelled and the fluorescent signal can be detected by exposing each cell individually to laser light as they pass through a small aperture, the essence of flow cytometry.
For flow cytometry to be beneficial in this project, we needed to purchase five new flow cytometers that could meet exacting standards. They needed to be sufficiently sensitive to identify rare cell populations, often with low levels of surface marker expression. For multicentre research projects, reproducibility of data between sites is essential. Hence, we needed excellent inter-machine reproducibility and the Figure 2. Initial training course with recently appointed flow managers (Credit: Dr Karen Hogg, University of York) | 10 manufacturer had to be able to provide service support across the region. In our search for the right flow cytometer to support the consortium, we settled upon the CytoFLEX, Beckman Coulter Life Sciences’ research flow cytometer, which uses avalanche photodiode detection to arrive at the required level of sensitivity. With assistance from Beckman Coulter, we devised and have run initial training courses with a group of recently appointed flow managers from each partner country, to share standard operating procedures, develop high-level data analysis strategies as well as to provide instruction in routine instrument maintenance.
Beckman Coulter also provides another important aid to reducing errors in flow cytometry for multisite projects such as this, namely freeze-dried antibody cocktails (DURAClone panels) [10], that allow highly multiplexed phenotyping of small volumes of blood added directly to a single tube. Particularly for investigators in remote locations, the use of dry, preformulated reagents, rather than liquid (‘wet’) antibodies, removes the need for a cold chain. Equally importantly, staining of cells when manual mixing of 15 or 16 antibodies is required can introduce data inconsistencies when conducted by different individuals and at different locations.
Together, these innovations have allowed us to establish a new network for flow cytometry in East Africa that will allow us to identify and functionally characterize and identify the types of immune cells present in the blood during these devastating diseases. We will match this data with similar multiplexed techniques in pathology to compare blood immune cell profiles with those of cells found in the skin, to give a more complete picture of the host response to infection before and after treatment or vaccination.
Future Directions
As mentioned, we are currently in the midst of an efficacy trial of our therapeutic vaccine, ChAd63-KH. The technology we are using is similar to that being used by researchers at the university of Oxford to develop a coronavirus vaccine. In short, we introduce two genes from Leishmania parasites (KMP-11 and HASPB1) into a well-studied chimpanzee adenovirus (ChAd63 viral vector). After vaccination with this vaccine, host cells become infected with the virus and express the Leishmania proteins in a way that can be recognized efficiently by the immune system. We are particularly interested in how well this vaccine can generate T|cells to fight the infection.
With the first of our clinical objectives now well underway – the ongoing therapeutic clinical trial in patients with PKDL will be completed in mid-2021 – we have two additional goals. The next, funded by EDCTP, is to start a new clinical trial to determine whether the vaccine can prevent progression from VL to PKDL. And finally, we hope to develop a human challenge model of leishmaniasis to test the vaccine for its ability to protect against infection by different forms of parasite. This would open the way to the development of a cost-effective prophylactic vaccine to prevent these diseases occurring in vulnerable populations across the world.
Our research also has larger ambitions for the long term. Our East African partners are also linked together through their work on leishmaniasis in drug development, as members of the Leishmaniasis East Africa Platform group, established to help coordinate drug development activities in the region by the Drugs for Neglected Diseases Partnership. Central questions about why the disease varies between countries are being addressed, and the increased capacity for flow cytometry will additionally support patient monitoring during drug trials conducted by DNDi or other groups. Indeed, through the capacity building this project provides, we hope this project will extend its reach beyond leishmaniasis, providing muchneeded support for research on other neglected diseases of poverty that affect people in the region, including bacterial, fungal, other parasitic and viral diseases. By continuing to demonstrate the analytical power of flow cytometry and its role in helping design much-needed therapies, we hope to open up additional discovery research possibilities for colleagues in Africa and around the world.
The research described in this article is part of the EDCTP2 programme supported by the European Union (grant number RIA2016V-1640; PREV_PKDL; https://www.prevpkdl.eu). The author Paul Kaye PhD, FRCPath, FMedSci Hull York Medical School, University of York, York, UK E-mail: paul.kaye@york.ac.uk
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Expert opinions from Dr Heidi Mendoza There are many assessments to make when adding a new test to a lab’s collection. Dr Heidi Mendoza, acting consultant clinical biochemist at Raigmore Hospital, Inverness, UK, shares her experiences and observations of doing exactly that in both ordinary circumstances and during a pandemic, as well as having to contend with the geographic challenges imposed by the nature of life in the Scottish Highlands. Can you provide a little background about yourself and where you work, please?
I am a clinical biochemist based in Raigmore Hospital, which is a small hospital in the Scottish Highlands. In my current role I provide clinical advice and interpretation for biochemistry tests for general practitioner (GP) practices and three hospitals across the Highlands. Working in the Highlands is incredibly rewarding, but also very challenging! It can take between 2 and 6|hours to travel between hospitals and our patients may have to travel by plane or boat to be seen, with journey times of +12|hours depending on where they live. It really puts the laboratories under pressure to get it right for the patient. Repeat testing isn’t as simple or straightforward as it would be in a city and we have to have excellent systems in place for reporting critical results and getting patients into hospital or transferring them between hospitals. Getting the right test, in the right place, with the right turnaround time is really important for our patients and for our clinicians. What are the usual circumstances in which you would think about bringing a new test into the lab’s repertoire?
Any new test is a cost pressure on our National Health Service (NHS) and can only be brought in when it demonstrates clear benefits for patients. We have brought in two new tests in the last 12|months that are good examples of the different ways we can bring in new tests to our laboratory.
The first test is the NT-proB-type natriuretic peptide (NTproBNP) test. NTproBNP is used to investigate patients with suspected heart failure and the results can be used to determine whether a patient needs an echocardiogram (ECHO) or not. If they do need an ECHO the NTproBNP result can be used to split patients into those who need urgent ECHO (2|weeks) or routine ECHO (6|weeks). In theory this is a perfect test to implement as it will benefit patients and is cost-effective with respect to the more expensive ECHO investigation. However, NTproBNP has been implemented in other hospitals without reducing ECHO waiting times or the number of ECHOs performed! To ensure that this didn’t happen in our service, I spent 6|months before implementation of the test liaising with cardiologists and GP representatives from across the Highland region. We changed the ECHO referral pathway to include NTproBNP and created useful guidance for GPs on when to, and importantly when not to, request NTproBNP. We implemented the test just under 1|year ago and have seen a positive effect on ECHO referrals. We will still have to attend a 1|year post-implementation review with the Hospital Board to present our audit data and show that investment in the service by introducing a new test has benefited patients and other areas of the service.
Procalcitonin is the second example. Procalcitonin is a test that can be used in the investigation of sepsis and guide the use of antibiotics. Procalcitonin was not a test available in our hospital before the COVID-19 pandemic. Procalcitonin is not increased in the majority of adult patients with COVID-19; however, an elevated procalcitonin may suggest superimposed bacterial infection and be used to guide treatment of these patients and improve patient outcomes. Early in the COVID-19 pandemic we were approached by our Intensive Care Unit (ITU) and Microbiology consultants who requested that procalcitonin be available for our COVID-19 patients in ITU to guide their antibiotic treatment. We implemented procalcitonin in less than 4|weeks with help from our instrument manufacturer, external quality assessment providers and other Scottish hospitals who provided anonymized patient serum with known values so that we could verify our assay as quickly as possible. We are now in the process of putting together a business case and following the evidence base which will determine whether we continue to offer the procalcitonin test. How would you usually go about adopting a new test?
As highlighted in the two examples above, we must agree a clinical need for a test and then liaise with the users of the service to find out how the test should be implemented into the patient-care pathway. Once we have worked out the clinical utility of the test, then we can carry out the laboratory verification of the test and the laboratory workflow. Verification is very straightforward. For example, the between-batch and within-batch precision, accuracy, linearity on dilution, interferences and sample stability for a test need to be evaluated. The implementation of the test then must be followed by an audit which shows that the test is being used as intended and giving the benefits predicted. If not, the test may need to be withdrawn. The hardest part of the entire process is agreeing how a test is going to be used and fitting it in to the patient-care pathway. In the situation of the COVID-19 pandemic, we have a new disease, caused by a new virus, and new tests that have been created very quickly. How do you start to use a new test in these circumstances – are there any differences in procedure?
There is no difference in the steps that need to be performed we just need to be able to do everything in a much shorter time frame. That is actually much easier than it sounds. In the NHS, the laboratories from different parts of the country are great about helping other laboratories. We regularly share protocols, data and learning. If a new test is released we’ll contact another laboratory and they’ll share their local experience and any problems they have had with the test.
For procalcitonin implementation I contacted the laboratory in Dundee, UK, and they helped us out by lending us kits and reagents, sending us anonymized patient serum with known procalcitonin values, and sharing their data and verification protocols. This allowed us to complete verification incredibly quickly. We will still have to gather the data and evaluate whether the test is providing the benefit that we predicted when we established the clinical need. What are the challenges regarding validation, reference levels, results interpretation and reporting?
Verifying tests is straightforward as we are always evaluating tests in clinical laboratories so are very experienced. Results interpretation can be quite difficult. If we need clinicians to change patient management based on a result then we have to provide them with very clear local guidance on what we want them to do with a result. This might be different from the action they would take in another hospital with different patient pathways, different pressures on patient turnaround times, and different diagnostic facilities. This is where good working relationships with users of the service are key to test implementation. If you just implement a new test without working out where it fits in the patient pathway, it doesn’t matter how great the test is, as it is unlikely to be used well and may not improve patient care. What do you have to think about in terms of logistics?
Many laboratories are understaffed due to a combination of unfilled vacancies and staff on long-term absence. The additional work involved in verifying and implementing a new test does put pressure on staff. However, NHS laboratory staff are highly trained and dedicated. When the staff know how a test is going to be used and the benefit to the local community, they support the implementation and the extra work involved.
Biocontainment and staff safety have been important considerations during the COVID-19 pandemic. We had to adhere to government guidance in the transport, analysis and disposal of samples from patients with suspected COVID-19. This changed laboratory workflows and slowed us down, creating longer turnaround times.
Logistics are a serious consideration for us owing to our geography. Reagent shortages or delays in deliveries have a big impact on small laboratories as they can’t store much surplus reagent stocks because of expiry dates. Unexpected overuse or underuse of a new test can be quite challenging and leave the laboratory short of tests or with expired, wasted kits. There are also several times during the year when the roads are impassable between our central and rural laboratories. We have been down to single numbers of tests remaining several times over the last few years or had failed delivery from manufacturers in winter. There was also a shortage of procalcitonin reagent as there was such a surge in the use of the test during the COVID-19 pandemic. Again, working closely with users of our laboratory services has enabled us to rationalize the use of the test until the global shortage of reagent ended. On a number of occasions we have also shared reagents with other Scottish laboratories to ensure that none of the laboratories were left without reagents. What has been learnt from the current coronavirus situation about diagnostic testing during a pandemic that would help to improve the process in future?
The coronavirus pandemic has shown how robust the infrastructure of the NHS is in Scotland and how adaptable laboratories can be when required. The laboratories really pulled together and worked towards a common goal delivering testing to COVID patients and non-COVID patients during a crisis. The two things that made this possible were: (1) Having a very clear goal – delivery of a service with new testing during a pandemic; and (2) Finances changes which needed to be made to deliver the service got rapid financial approval. How do we take these lessons learned and apply it to the routine delivery of laboratory services? Finance will always be a limiting factor – as it should be! Healthcare is expensive and it is up to us as healthcare professionals to deliver a cost-effective and affordable service. In contrast, having a clear goal, is definitely something that we could do better in the future. In the case of the pandemic, laboratories found different solutions based on local geography, resources and incidence of COVID. The changes made by laboratories in the remote Highlands and Islands were similar, but different than those made by laboratories in major cities. The staff that delivered the service found the best solutions to the goals set by the government – that is the real lesson we need to take away. We need to give very clear goals to services and let local expertise and knowledge drive the changes to solve the problem. The expert Heidi Mendoza BSc MSc PhD RCPath Blood Sciences Department, Raigmore Hospital, Inverness IV2 3UJ, UK E-mail: heidi.mendoza@nhs.net
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LGC has acquired The Native Antigen Company (NAC), one of the world’s leading suppliers of high quality infectious disease antigens and antibodies.
NAC is a developer, manufacturer and supplier of critical reagents to the in vitro diagnostic (IVD), pharmaceutical and academic sectors. It offers a comprehensive portfolio of native and recombinant infectious disease antigens and related products including pathogen receptors, virus-like particles and antibodies for use in immunoassay applications, vaccine development and quality control solutions. NAC was one of the first companies globally to offer antigens for SARS-COV-2 and continues to play an important role in supporting the global response to the COVID-19 pandemic.
The acquisition strengthens LGC’s existing product offering to the IVD sector, which includes a range of quality assurance tools, immunoassay reagents and disease state plasma as well as probes and primers for molecular diagnostics.
“NAC is a natural fit with our clinical diagnostics business and will enable us to provide an expanded portfolio of critical reagents to our customers. NAC’s focus on infectious disease is highly complementary with our existing offer to this segment comprising controls, reference materials, MDx tools and other components,” said Michael Sweatt, Executive Vice President and General Manager, Clinical Diagnostics, LGC.
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Avacta Group, the developer of Affimer biotherapeutics and reagents, has entered into a collaboration with Integumen to evaluate recently generated Affimer reagents that bind the SARS-COV-2 spike protein for the detection of the coronavirus in waste water, to provide a real-time alert system to warn of localised COVID-19 outbreaks.
Over 60 percent of COVID-19 positive patients had gastrointestinal symptoms, such as diarrhoea, nausea and vomiting, and the SARS-COV-2 virus was found in their faecal samples. Sampling waste water from households may therefore provide an early warning system for localised outbreaks in communities.
Recently, Avacta announced that it had generated a number of highly specific Affimer reagents that detect the SARS-COV-2 virus spike protein for use in diagnostic tests and in neutralising therapies.
The collaboration with Integumen, announced 13 July, aims to evaluate some of these Affimer reagents in next-generation sensors, based on the real-time bacteria detection and alert system1 developed by Rinocloud, a subsidiary of Integumen, with the aim of integrating these sensors into Modern Water’s Microtox water contamination system to detect the coronavirus. The award-winning Microtox system, which can detect the presence of contaminating bacteria, virus and toxins, is distributed by Modern Water and has a global footprint of over 3,000 installations. The proposed Affimer sensors would be consumable items to be replaced on a roughly monthly basis.
Once initial testing of the Affimer reagents is completed over the next few weeks, validation of the sensors will be carried out using SARS-COV-2 virus samples in a containment level 3 laboratory at the University of Aberdeen. Upon successful completion of this evaluation, Integumen and Avacta will enter into a supply agreement to allow Integumen to manufacture and commercialise the waste water detection sensors globally by retrofitting into Microtox systems.
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Byondis B.V. (formerly Synthon Biopharmaceuticals) announced that Quantum Leap Healthcare Collaborative (Quantum Leap) selected the company’s investigational antibody-drug conjugate (ADC) SYD985 ([vic-]trastuzumab duocarmazine) for a new investigational treatment arm in its ongoing I-SPY 2 TRIAL for neoadjuvant treatment of locally advanced breast cancer. This treatment arm will focus on treatment for HER2-low early-stage breast cancer.
The I-SPY 2 TRIAL (Investigation of Serial studies to Predict Your Therapeutic Response with Imaging And moLecular analysis) is a standing Phase II randomized, controlled, multicentre study aimed at rapidly screening and identifying promising treatments in specific subgroups of women with newly-diagnosed, high-risk, locally advanced breast cancer (Stage II/III). Quantum Leap, sponsor of the I-SPY 2 TRIAL, leads a pre-competitive consortium that includes the U.S. Food & Drug Administration (FDA), industry, patient advocates, philanthropic sponsors, and clinicians from 16 major U.S. cancer research centres.
The new I-SPY 2 treatment arm will evaluate SYD985 against standard of care therapy in Stage II/III early-stage, high-risk breast cancer patients, with a focus on tumours with heterogeneous and low HER2 expression. Byondis will supply the investigational drug and provide financial and regulatory support. Quantum Leap, as sponsor, will provide the clinical sites and clinical expertise.
SYD985 is Byondis’ most advanced ADC, targeting a range of HER2-positive cancers such as metastatic breast cancer (MBC) and endometrial cancer. The company is currently conducting a Phase III study of SYD985 (TULIP or SYD985.002) to compare its efficacy and safety to physician’s choice treatment in patients with HER2-positive unresectable locally advanced or metastatic breast cancer. Previously, the FDA granted fast track designation for SYD985 based on promising data from heavily pre-treated last-line HER2-positive MBC patients participating in a two-part Phase I clinical trial (SYD985.001).
SYD985 uses Byondis’ unique, proprietary linker-drug (LD) technology. Although marketed ADCs have improved therapeutic indices compared to classical non-targeted chemotherapeutic agents, there is still room for improvement.
SYD985 is comprised of the monoclonal antibody trastuzumab and a cleavable linker-drug called valine-citrulline-seco-DUocarmycin-hydroxyBenzamide-Azaindole (vc-seco-DUBA). The antibody part of SYD985 binds to HER2 on the surface of the cancer cell and the ADC is internalized by the cell. After proteolytic cleavage of the linker, the inactive cytotoxin is activated and DNA damage is induced, resulting in tumour cell death. SYD985 can be considered a form of targeted chemotherapy.
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Dante Labs, a pioneer and leader in genomic testing, Cambridge Cancer Genomics (CCG.ai), a software developer specialising in data-driven precision oncology, and Nonacus, a provider of genetic testing products for precision medicine and liquid biopsy, have signed a collaboration agreement.
In a joint statement they said the partnership aims to build the most comprehensive and patient-centric tumour profiling service enabling improved cancer patient management, treatment and monitoring. By combining Dante Labs’ experience and capacity in delivering a sequencing service for both solid tumour and cell free circulating tumour DNA from liquid biopsies, Nonacus’ sensitive targeted pan-cancer NGS libraries, and CCG.ai’s industry leading AI powered software platform, OncOS, the companies will enable precision oncology at scale.
Improving outcomes for cancer patients means ensuring they have the right drug, at the right time to beat their cancer. This means understanding the molecular profile of the individual cancer and using that data to recommend treatments or clinical trials. Oncologists and clinical researchers will be able to send samples for processing to Dante Labs, who will use library preparation kits from Nonacus and software from CCG.ai to create a sample to report solution. If there are actionable mutations, the report will recommend the right treatments for those mutations, if there are novel or unactionable mutations, the software will also be able to match possible clinical trials. Chris Sale, CEO of Nonacus, said: “Long turn-around time and lack of clinically oriented analysis are the main obstacles to fully deliver the potential of cancer genomics to patients. This partnership will provide the flexibility and accuracy that oncology professionals need to integrate cancer genomics into the care of their patients. The COVID pandemic has increased the backlog of genetic testing for cancer, potentially leaving many suspected cancers unconfirmed and treatments delayed. Dante Labs are one of the biggest clinical sequencing hub in Europe able to process large numbers of samples in high throughput. It is our hope that by combining AI software from CCG.ai and our library preparation kits, together we will be able to process samples and provide bioinformatic analysis critical to determining the best treatment path for patients.”
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Start Codon, a new model of life science and healthcare business accelerator, has announced its first cohort of start-up companies. Start Codon aims to minimise risk and translate early stage research into successful start-ups, ready for funding and partnership. Start Codon has worked closely with four life science and healthcare companies that were enrolled into the programme in February this year.
They are:
Enhanc3D Genomics, a functional genomics spin-out of the Babraham Institute (Cambridge, UK) whose platform technology links non-coding sequence variants to their target genes in order to identify novel therapeutic targets
Drishti Discoveries, a start-up leveraging a proprietary gene silencing technology to develop therapies for rare inherited diseases
Spirea, a spin-out from the University of Cambridge, who is developing the next generation of antibody drug conjugate cancer therapeutics which carry more drug payload to tumour cells, resulting in greater efficacy, tolerability and the ability to treat more cancer patients
Semarion, a University of Cambridge spin-out, who is revolutionising cell-based assays for drug discovery and life science through its proprietary SemaCyte microcarrier platform, which leverages novel materials physics for assay miniaturisation, multiplexing, and automation
Start Codon plans to invest in and support up to 50 start-up companies over the next five years. The accelerator is now accepting applications for its second and third cohorts of companies. Early stage start-up companies in the life sciences and healthcare space are invited to apply via https://startcodon.co/application-form
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The ReactoMate DATUM from Asynt is a high quality, dual-rod stainless steel and aluminium support system built to ensure the stability and safety of your lab reactor. Sturdy, yet compact, the ReactoMate DATUM support system can accommodate a wide range of reaction vessels from 100 mL up to 5000 mL.
Designed with user-friendliness in mind, the ReactoMate DATUM support system incorporates a suite of innovative features.
Changing a vessel supported by the ReactoMate DATUM is as simple as “Clip & Click”. The novel neck clamp allows fast changeover between reactor vessel sizes thereby enabling simple reaction scale-up, whilst the ingenious mounting mechanism ensures excellent stability and alignment every time.
The Reactomate DATUM support system is fully compatible with all leading brands of overhead stirrers and circulator heating/cooling systems. Designed by chemists for chemists, low-friction polymer bearings line both the overhead stirrer alignment chuck and the neck support to ensure smooth and easy operation.
Ideally suited for use within a benchtop fume hood, adjustable feet allow you to level the ReactoMate DATUM support system ensuring stability and security while you work. Each DATUM system is also supplied with a moulded drip tray that fits perfectly within the base of the support, for safely catching any drips and spills from the reaction vessel during draining.
With a wide range of accessories and upgrades available, including drain manifolds and automation packages, the ReactoMate DATUM support system is the perfect all-rounder for laboratory scale reactions.
For more information, visit: www.asynt.com/product/reactomate-datum
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Concarlo Holdings has received a US patent for IpY, a novel therapeutic peptide that addresses drug-resistant breast cancer by targeting a unique cellular pathway — p27Kip1. The patent is the latest step in Concarlo’s journey to commercialize revolutionary medicines for metastatic breast cancer.
Concarlo has also announced that a new provisional patent application has been filed for modified versions of the therapeutic peptide that are believed to exhibit enhanced bioavailability. Concarlo is a Brooklyn, New York-based biotechnology innovator dedicated to developing sophisticated, targeted therapies and diagnostics in the oncology space. The IpY technology is the first to address the high incidence of drug-refractory disease that develops with currently available CDK4 inhibitor (CDK4i) treatments. Such a solution has the potential to drastically increase overall survival of breast cancer patients.
The recent introduction of CDK4i drugs, a class of medicines that directly targets the CDK4/6 pathway implicated in many malignancies, has had a significant impact on the way in which metastatic breast cancer is managed. However, such therapeutics are associated with patients transitioning to a treatment-resistant form of the condition, despite initial extended periods of remission. Backed by more than 20 years of research and development expertise, Concarlo has developed IpY and a companion diagnostic, ApY, to effectively overcome the issue of CDK4i resistance and roll out a more targeted treatment approach for optimized patient outcomes.
“Despite the clinical efficacy of CDK4 inhibitors, we’re seeing that primary or secondary resistance to therapy is presenting a significant challenge to overall survival,” said Dr. Dominique Bridon, Chief Development Officer at Concarlo. “With the IpY technology and its unique mechanism of action, we’re effectively targeting CDK4 while simultaneously inhibiting another target — CDK2 — which has been found to be a key molecular player in the development of drug resistance. In doing so, we are the first company to successfully address the CDK4i resistance issue to provide long-term durable tumour arrest. Combined with its highly specific targeting and low toxicity profile, the positive impact of this drug on the breast cancer treatment landscape is hard to understate.”
Concarlo was formed in 2016 and is supported by a team of internationally renowned experts forming its Scientific Advisory Board. To date, the company has raised more than $3.1 million to support the development, improvement, and commercialization of its IpY and ApY technologies to bring a precision medicine approach to breast cancer management. The newly issued patent for IpY and the provisional patent application for modified versions of the peptide are the first key milestones in Concarlo’s plan to build an extensive patent estate to maintain market exclusivity for its clinically relevant therapeutics.
https://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png003wmediahttps://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png3wmedia2020-08-26 09:31:372021-01-08 11:07:46Concarlo awarded patent for novel therapeutic peptide for drug-resistant breast cancer
Sphere Fluidics, a company developing single cell analysis systems underpinned by its patented picodroplet technology, and Heriot-Watt University, a specialist, pioneering Scottish University, have been awarded a Knowledge Transfer Partnership (KTP) grant from Innovate UK, the UK’s innovation agency. The grant will facilitate the development of novel droplet generator instrumentation, which will be used to expand Sphere Fluidics’ portfolio of microfluidic instruments for advanced biologics discovery and therapeutic cell line development.
Awarded to promote the collaboration of knowledge, technology and skills within the UK Knowledge Base, the KTP has been granted to Sphere Fluidics, in partnership with Dr. Graeme Whyte, Associate Professor at Heriot-Watt University. The two-year project will develop next-generation intelligent instrumentation and advance research across a range of picodroplet techniques, allowing scientists to discover rare cell phenotypes and to help to solve a range of biological questions ranging from antibody discovery to antimicrobial resistance, enzyme evolution and synthetic biology. The novel platform for semi-automated picodroplet production will be employed by the company to improve control of droplet production, using advanced imaging technology.
As part of the project, Dr. John McGrath has been appointed to Sphere Fluidics’ team as a Research Scientist in physics and engineering, to support the transfer of cutting-edge research into the company’s portfolio of single-cell analysis instruments, including for several new commercial products.
Dr. Marian Rehak, VP of Research and Development at Sphere Fluidics, said: “This innovative project with Heriot-Watt University, will bring together aspects of microfluidic and optical design, technology development and product design engineering to develop a new class of instrument for cell-based picodroplet discovery. We are delighted to have been awarded the KTP Fellowship and to welcome Dr. John McGrath to the Sphere Fluidics team. The work demonstrates the importance of collaboration between academic and industrial partners to support the advancement of novel microfluidic technologies for ground-breaking research.”
Dr. John McGrath, Research Scientist at Sphere Fluidics, commented: “I am thrilled to be working alongside commercial and academic leaders in the research and development of microfluidic instruments and technology. The ease of use and broader application set of the instrument to be developed in this project should lower the barrier to entry for a wide number of scientists, who are focused upon high-throughput screening, synthetic biology, gene editing, and antimicrobial resistance workflows. The technology has the potential to be a key driver in increasing the uptake of picodroplet microfluidic instruments.”
https://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png003wmediahttps://clinlabint.com/wp-content/uploads/sites/2/2020/06/clinlab-logo.png3wmedia2020-08-26 09:31:372021-01-08 11:07:46Sphere Fluidics, Heriot-Watt University collaborate to develop next generation droplet generator instrumentation
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Flow cytometry: a critical technique in combating leishmaniasis
, /in E-News /by 3wmediaby Professor Paul Kaye
Leishmaniasis is classified as a neglected tropical disease. It is the cause of a huge health burden and is common in Asia, Africa, South and Central America, and even southern Europe. This article discusses how flow cytometry can help to evaluate diagnosis, monitor the effects of therapy and help in the creation of a vaccine.
Background
The leishmaniases are a family of devastating diseases, affecting a great many people across the globe and presenting a significant risk to both public health and socioeconomic development. The leishmaniases are vector-borne diseases, caused by infection with one of 20 species of the parasitic protozoan Leishmania (Fig. 1), transmitted through the bite of the infected female phlebotomine sand fly.
They can be broadly classified as tegumentary leishmaniases (TLs), affecting the skin and mucosa, and visceral leishmaniasis (VL), affecting internal organs. Whereas VL is responsible for over 20¦000 deaths per year, TL are non-life-threatening, chronic and potentially disfiguring, and account for around two-thirds of the global disease burden.
Within TL, there are three subtypes: self-healing lesions at the location of sand fly bite (cutaneous leishmaniasis; CL), lesions that spread from the original skin lesion to the mucosae (mucosal leishmaniasis; ML), and those which spread uncontrolled across the body (disseminated or diffuse cutaneous leishmaniasis; DCL). VL, also known as kala azar, involves major organs including the spleen, liver and bone marrow. In addition, patients recovering from VL after drug treatment often develop post kala-azar dermal leishmaniasis (PKDL), a chronic skin condition, characterized by nodular or macular lesions beginning on the face and spreading to the trunk and arms. As it may develop in up to half of patients previously treated and apparently cured from VL, it is thought that PKDL plays a central role in community transmission of VL.
The World Health Organization designates leishmaniasis as a neglected tropical disease (NTD), which together affect more than one|billion people across 149 countries worldwide; true prevalence may be even higher. Disproportionately, NTDs affect the poorest, malnourished individuals, and contribute to a vicious circle of poverty and disease. The significant physical marks, including ulcers, often left in the wake of the TLs may have an impact on mental health and perpetuate social stigma associated with the diseases [5]. There are over 1|million new cases of TL and 0.5|million new cases of VL each year, which together account for the loss of approximately 2.4|million disability-adjusted life years.
Treatment challenges
Leishmaniasis treatment can be quite difficult since at-risk populations may lack access to healthcare, and the limited battery of drugs has been increasingly compromised by resistance. Additionally, because the parasites in question are eukaryotic, they are not dissimilar from human cells, so the medication is also liable to be harmful – even fatal – to host as well as to pathogen.
Although the burden of VL in South Asia has been reduced with single-dose liposomal amphotericin B, the drug is less effective in other geographic locations, namely East Africa. Various drug combinations have been tested, unsuccessfully, and new chemical entities and immune-modulators are in early stages of development and as yet untested in the field. Unfortunately, little has changed in the treatment for CL for the past 50|years.
No vaccines are currently approved for any form of human leishmaniasis, although vaccines for canine VL have reached the market. Barriers to vaccine development include the limited investment in leishmaniases R&D and the high costs involved in bringing new products to those that need them.
Current work
My work on leishmaniasis has taken a holistic view, rooted in the immunology of the host-parasite interaction, but employing tools and approaches that span many disciplines: mathematics, ecology, vector biology and most recently neuroscience. Thirty years of discovery science has led to the development of a candidate for a therapeutic vaccine for PKDL, the mysterious sequela to VL [6]. ‘Therapeutic’ vaccines are given after an individual is infected with a pathogen and are designed to enhance our immune system and help eliminate the infection.
With colleagues from Sudan, we are in the midst of a phase IIb clinical trial funded by the Wellcome Trust, evaluating the efficacy of this therapeutic vaccine in Sudanese patients with persistent PKDL.
However, the research has been a long time in the making and has a long way to go. To continue to make progress, we linked with colleagues in Ethiopia, Kenya and Uganda and at the European Vaccine Initiative (http://www.euvaccine.eu/) in Germany, to develop a new research consortium to evaluate the immune status of people suffering from leishmaniasis. For example, using flow cytometry for blood and multiplexed immunohistochemistry for tissue biopsies, we can enumerate the proportions of lymphocytes, monocytes and neutrophils based on surface marker expression (e.g. CD3, CD19, CD14, CD16), and characterize their function, for instance by expression of cytokines (e.g. interferon-gamma) or other cell surface proteins that define function state. To support this endeavour, we recently received a grant from the European & Developing Countries Clinical Trials Partnership (EDCTP) that will allow us to not only extend our vaccine programme in Sudan [9] but also to address other important research challenges.
To develop vaccines and indeed new drugs, we often need tools capable of performing in-depth comparisons of how the body’s immune system is coping with the infection when a patient is first admitted to hospital and how it changes as the patient undergoes treatment and is hopefully cured. For example, recent evidence suggests that during infection, T lymphocytes may become ‘exhausted’ and unable to fight infection and the exhausted state can be identified by expression of surface molecules such as programmed cell death protein|1 (PD-1) and lymphocyte activation gene 3 protein (LAG-3). It is important to know if exhaustion can be reversed following treatment or whether we need to stimulate new populations of T lymphocytes. By understanding these nuanced changes in immune cells in our blood, we can design ways to improve how vaccines and drugs work in concert with immune cells, and understand why some patients might relapse from their disease or develop PKDL. Flow cytometry is a central tool for immunologists and plays a critical role in uncovering mechanisms of immunity and in assessing how well vaccines work and biomarkers of drug response. It uses antibodies that recognize specific molecules or markers on the surface or inside immune cells, such as those mentioned above, that help us predict their function. These antibodies are fluorescently labelled and the fluorescent signal can be detected by exposing each cell individually to laser light as they pass through a small aperture, the essence of flow cytometry.
For flow cytometry to be beneficial in this project, we needed to purchase five new flow cytometers that could meet exacting standards. They needed to be sufficiently sensitive to identify rare cell populations, often with low levels of surface marker expression. For multicentre research projects, reproducibility of data between sites is essential. Hence, we needed excellent inter-machine reproducibility and the Figure 2. Initial training course with recently appointed flow managers (Credit: Dr Karen Hogg, University of York) | 10 manufacturer had to be able to provide service support across the region. In our search for the right flow cytometer to support the consortium, we settled upon the CytoFLEX, Beckman Coulter Life Sciences’ research flow cytometer, which uses avalanche photodiode detection to arrive at the required level of sensitivity. With assistance from Beckman Coulter, we devised and have run initial training courses with a group of recently appointed flow managers from each partner country, to share standard operating procedures, develop high-level data analysis strategies as well as to provide instruction in routine instrument maintenance.
Beckman Coulter also provides another important aid to reducing errors in flow cytometry for multisite projects such as this, namely freeze-dried antibody cocktails (DURAClone panels) [10], that allow highly multiplexed phenotyping of small volumes of blood added directly to a single tube. Particularly for investigators in remote locations, the use of dry, preformulated reagents, rather than liquid (‘wet’) antibodies, removes the need for a cold chain. Equally importantly, staining of cells when manual mixing of 15 or 16 antibodies is required can introduce data inconsistencies when conducted by different individuals and at different locations.
Together, these innovations have allowed us to establish a new network for flow cytometry in East Africa that will allow us to identify and functionally characterize and identify the types of immune cells present in the blood during these devastating diseases. We will match this data with similar multiplexed techniques in pathology to compare blood immune cell profiles with those of cells found in the skin, to give a more complete picture of the host response to infection before and after treatment or vaccination.
Future Directions
As mentioned, we are currently in the midst of an efficacy trial of our therapeutic vaccine, ChAd63-KH. The technology we are using is similar to that being used by researchers at the university of Oxford to develop a coronavirus vaccine. In short, we introduce two genes from Leishmania parasites (KMP-11 and HASPB1) into a well-studied chimpanzee adenovirus (ChAd63 viral vector). After vaccination with this vaccine, host cells become infected with the virus and express the Leishmania proteins in a way that can be recognized efficiently by the immune system. We are particularly interested in how well this vaccine can generate T|cells to fight the infection.
With the first of our clinical objectives now well underway – the ongoing therapeutic clinical trial in patients with PKDL will be completed in mid-2021 – we have two additional goals. The next, funded by EDCTP, is to start a new clinical trial to determine whether the vaccine can prevent progression from VL to PKDL. And finally, we hope to develop a human challenge model of leishmaniasis to test the vaccine for its ability to protect against infection by different forms of parasite. This would open the way to the development of a cost-effective prophylactic vaccine to prevent these diseases occurring in vulnerable populations across the world.
Our research also has larger ambitions for the long term. Our East African partners are also linked together through their work on leishmaniasis in drug development, as members of the Leishmaniasis East Africa Platform group, established to help coordinate drug development activities in the region by the Drugs for Neglected Diseases Partnership. Central questions about why the disease varies between countries are being addressed, and the increased capacity for flow cytometry will additionally support patient monitoring during drug trials conducted by DNDi or other groups. Indeed, through the capacity building this project provides, we hope this project will extend its reach beyond leishmaniasis, providing muchneeded support for research on other neglected diseases of poverty that affect people in the region, including bacterial, fungal, other parasitic and viral diseases. By continuing to demonstrate the analytical power of flow cytometry and its role in helping design much-needed therapies, we hope to open up additional discovery research possibilities for colleagues in Africa and around the world.
The research described in this article is part of the EDCTP2 programme supported by the European Union (grant number RIA2016V-1640; PREV_PKDL; https://www.prevpkdl.eu).
The author
Paul Kaye PhD, FRCPath, FMedSci
Hull York Medical School, University of York, York, UK
E-mail: paul.kaye@york.ac.uk
Introducing new tests to a laboratory’s repertoire
, /in Corona News, E-News /by 3wmediaExpert opinions from Dr Heidi Mendoza
There are many assessments to make when adding a new test to a lab’s collection. Dr Heidi Mendoza, acting consultant clinical biochemist at Raigmore Hospital, Inverness, UK, shares her experiences and observations of doing exactly that in both ordinary circumstances and during a pandemic, as well as having to contend with the geographic challenges imposed by the nature of life in the Scottish Highlands.
Can you provide a little background about yourself and where you work, please?
I am a clinical biochemist based in Raigmore Hospital, which is a small hospital in the Scottish Highlands. In my current role I provide clinical advice and interpretation for biochemistry tests for general practitioner (GP) practices and three hospitals across the Highlands. Working in the Highlands is incredibly rewarding, but also very challenging! It can take between 2 and 6|hours to travel between hospitals and our patients may have to travel by plane or boat to be seen, with journey times of +12|hours depending on where they live. It really puts the laboratories under pressure to get it right for the patient. Repeat testing isn’t as simple or straightforward as it would be in a city and we have to have excellent systems in place for reporting critical results and getting patients into hospital or transferring them between hospitals. Getting the right test, in the right place, with the right turnaround time is really important for our patients and for our clinicians.
What are the usual circumstances in which you would think about bringing a new test into the lab’s repertoire?
Any new test is a cost pressure on our National Health Service (NHS) and can only be brought in when it demonstrates clear benefits for patients. We have brought in two new tests in the last 12|months that are good examples of the different ways we can bring in new tests to our laboratory.
The first test is the NT-proB-type natriuretic peptide (NTproBNP) test. NTproBNP is used to investigate patients with suspected heart failure and the results can be used to determine whether a patient needs an echocardiogram (ECHO) or not. If they do need an ECHO the NTproBNP result can be used to split patients into those who need urgent ECHO (2|weeks) or routine ECHO (6|weeks). In theory this is a perfect test to implement as it will benefit patients and is cost-effective with respect to the more expensive ECHO investigation. However, NTproBNP has been implemented in other hospitals without reducing ECHO waiting times or the number of ECHOs performed! To ensure that this didn’t happen in our service, I spent 6|months before implementation of the test liaising with cardiologists and GP representatives from across the Highland region. We changed the ECHO referral pathway to include NTproBNP and created useful guidance for GPs on when to, and importantly when not to, request NTproBNP. We implemented the test just under 1|year ago and have seen a positive effect on ECHO referrals. We will still have to attend a 1|year post-implementation review with the Hospital Board to present our audit data and show that investment in the service by introducing a new test has benefited patients and other areas of the service.
Procalcitonin is the second example. Procalcitonin is a test that can be used in the investigation of sepsis and guide the use of antibiotics. Procalcitonin was not a test available in our hospital before the COVID-19 pandemic. Procalcitonin is not increased in the majority of adult patients with COVID-19; however, an elevated procalcitonin may suggest superimposed bacterial infection and be used to guide treatment of these patients and improve patient outcomes. Early in the COVID-19 pandemic we were approached by our Intensive Care Unit (ITU) and Microbiology consultants who requested that procalcitonin be available for our COVID-19 patients in ITU to guide their antibiotic treatment. We implemented procalcitonin in less than 4|weeks with help from our instrument manufacturer, external quality assessment providers and other Scottish hospitals who provided anonymized patient serum with known values so that we could verify our assay as quickly as possible. We are now in the process of putting together a business case and following the evidence base which will determine whether we continue to offer the procalcitonin test.
How would you usually go about adopting a new test?
As highlighted in the two examples above, we must agree a clinical need for a test and then liaise with the users of the service to find out how the test should be implemented into the patient-care pathway. Once we have worked out the clinical utility of the test, then we can carry out the laboratory verification of the test and the laboratory workflow. Verification is very straightforward. For example, the between-batch and within-batch precision, accuracy, linearity on dilution, interferences and sample stability for a test need to be evaluated. The implementation of the test then must be followed by an audit which shows that the test is being used as intended and giving the benefits predicted. If not, the test may need to be withdrawn. The hardest part of the entire process is agreeing how a test is going to be used and fitting it in to the patient-care pathway.
In the situation of the COVID-19 pandemic, we have a new disease, caused by a new virus, and new tests that have been created very quickly. How do you start to use a new test in these circumstances – are there any differences in procedure?
There is no difference in the steps that need to be performed we just need to be able to do everything in a much shorter time frame. That is actually much easier than it sounds. In the NHS, the laboratories from different parts of the country are great about helping other laboratories. We regularly share protocols, data and learning. If a new test is released we’ll contact another laboratory and they’ll share their local experience and any problems they have had with the test.
For procalcitonin implementation I contacted the laboratory in Dundee, UK, and they helped us out by lending us kits and reagents, sending us anonymized patient serum with known procalcitonin values, and sharing their data and verification protocols. This allowed us to complete verification incredibly quickly. We will still have to gather the data and evaluate whether the test is providing the benefit that we predicted when we established the clinical need.
What are the challenges regarding validation, reference levels, results interpretation and reporting?
Verifying tests is straightforward as we are always evaluating tests in clinical laboratories so are very experienced. Results interpretation can be quite difficult. If we need clinicians to change patient management based on a result then we have to provide them with very clear local guidance on what we want them to do with a result. This might be different from the action they would take in another hospital with different patient pathways, different pressures on patient turnaround times, and different diagnostic facilities. This is where good working relationships with users of the service are key to test implementation. If you just implement a new test without working out where it fits in the patient pathway, it doesn’t matter how great the test is, as it is unlikely to be used well and may not improve patient care.
What do you have to think about in terms of logistics?
Many laboratories are understaffed due to a combination of unfilled vacancies and staff on long-term absence. The additional work involved in verifying and implementing a new test does put pressure on staff. However, NHS laboratory staff are highly trained and dedicated. When the staff know how a test is going to be used and the benefit to the local community, they support the implementation and the extra work involved.
Biocontainment and staff safety have been important considerations during the COVID-19 pandemic. We had to adhere to government guidance in the transport, analysis and disposal of samples from patients with suspected COVID-19. This changed laboratory workflows and slowed us down, creating longer turnaround times.
Logistics are a serious consideration for us owing to our geography. Reagent shortages or delays in deliveries have a big impact on small laboratories as they can’t store much surplus reagent stocks because of expiry dates. Unexpected overuse or underuse of a new test can be quite challenging and leave the laboratory short of tests or with expired, wasted kits. There are also several times during the year when the roads are impassable between our central and rural laboratories. We have been down to single numbers of tests remaining several times over the last few years or had failed delivery from manufacturers in winter. There was also a shortage of procalcitonin reagent as there was such a surge in the use of the test during the COVID-19 pandemic. Again, working closely with users of our laboratory services has enabled us to rationalize the use of the test until the global shortage of reagent ended. On a number of occasions we have also shared reagents with other Scottish laboratories to ensure that none of the laboratories were left without reagents.
What has been learnt from the current coronavirus situation about diagnostic testing during a pandemic that would help to improve the process in future?
The coronavirus pandemic has shown how robust the infrastructure of the NHS is in Scotland and how adaptable laboratories can be when required. The laboratories really pulled together and worked towards a common goal delivering testing to COVID patients and non-COVID patients during a crisis. The two things that made this possible were: (1) Having a very clear goal – delivery of a service with new testing during a pandemic; and (2) Finances changes which needed to be made to deliver the service got rapid financial approval. How do we take these lessons learned and apply it to the routine delivery of laboratory services? Finance will always be a limiting factor – as it should be! Healthcare is expensive and it is up to us as healthcare professionals to deliver a cost-effective and affordable service. In contrast, having a clear goal, is definitely something that we could do better in the future. In the case of the pandemic, laboratories found different solutions based on local geography, resources and incidence of COVID. The changes made by laboratories in the remote Highlands and Islands were similar, but different than those made by laboratories in major cities. The staff that delivered the service found the best solutions to the goals set by the government – that is the real lesson we need to take away. We need to give very clear goals to services and let local expertise and knowledge drive the changes to solve the problem.
The expert
Heidi Mendoza BSc MSc PhD RCPath
Blood Sciences Department, Raigmore Hospital, Inverness IV2 3UJ, UK
E-mail: heidi.mendoza@nhs.net
LGC acquires The Native Antigen Company
, /in Corona News, E-News /by 3wmediaLGC has acquired The Native Antigen Company (NAC), one of the world’s leading suppliers of high quality infectious disease antigens and antibodies.
NAC is a developer, manufacturer and supplier of critical reagents to the in vitro diagnostic (IVD), pharmaceutical and academic sectors. It offers a comprehensive portfolio of native and recombinant infectious disease antigens and related products including pathogen receptors, virus-like particles and antibodies for use in immunoassay applications, vaccine development and quality control solutions. NAC was one of the first companies globally to offer antigens for SARS-COV-2 and continues to play an important role in supporting the global response to the COVID-19 pandemic.
The acquisition strengthens LGC’s existing product offering to the IVD sector, which includes a range of quality assurance tools, immunoassay reagents and disease state plasma as well as probes and primers for molecular diagnostics.
“NAC is a natural fit with our clinical diagnostics business and will enable us to provide an expanded portfolio of critical reagents to our customers. NAC’s focus on infectious disease is highly complementary with our existing offer to this segment comprising controls, reference materials, MDx tools and other components,” said Michael Sweatt, Executive Vice President and General Manager, Clinical Diagnostics, LGC.
Avacta, Integumen collaborate for detection of SARS-COV-2 in waste water
, /in Corona News, E-News /by 3wmediaAvacta Group, the developer of Affimer biotherapeutics and reagents, has entered into a collaboration with Integumen to evaluate recently generated Affimer reagents that bind the SARS-COV-2 spike protein for the detection of the coronavirus in waste water, to provide a real-time alert system to warn of localised COVID-19 outbreaks.
Over 60 percent of COVID-19 positive patients had gastrointestinal symptoms, such as diarrhoea, nausea and vomiting, and the SARS-COV-2 virus was found in their faecal samples. Sampling waste water from households may therefore provide an early warning system for localised outbreaks in communities.
Recently, Avacta announced that it had generated a number of highly specific Affimer reagents that detect the SARS-COV-2 virus spike protein for use in diagnostic tests and in neutralising therapies.
The collaboration with Integumen, announced 13 July, aims to evaluate some of these Affimer reagents in next-generation sensors, based on the real-time bacteria detection and alert system1 developed by Rinocloud, a subsidiary of Integumen, with the aim of integrating these sensors into Modern Water’s Microtox water contamination system to detect the coronavirus. The award-winning Microtox system, which can detect the presence of contaminating bacteria, virus and toxins, is distributed by Modern Water and has a global footprint of over 3,000 installations. The proposed Affimer sensors would be consumable items to be replaced on a roughly monthly basis.
Once initial testing of the Affimer reagents is completed over the next few weeks, validation of the sensors will be carried out using SARS-COV-2 virus samples in a containment level 3 laboratory at the University of Aberdeen. Upon successful completion of this evaluation, Integumen and Avacta will enter into a supply agreement to allow Integumen to manufacture and commercialise the waste water detection sensors globally by retrofitting into Microtox systems.
Byondis investigational breast cancer drug selected for trial
, /in E-News /by 3wmediaByondis B.V. (formerly Synthon Biopharmaceuticals) announced that Quantum Leap Healthcare Collaborative (Quantum Leap) selected the company’s investigational antibody-drug conjugate (ADC) SYD985 ([vic-]trastuzumab duocarmazine) for a new investigational treatment arm in its ongoing I-SPY 2 TRIAL for neoadjuvant treatment of locally advanced breast cancer. This treatment arm will focus on treatment for HER2-low early-stage breast cancer.
The I-SPY 2 TRIAL (Investigation of Serial studies to Predict Your Therapeutic Response with Imaging And moLecular analysis) is a standing Phase II randomized, controlled, multicentre study aimed at rapidly screening and identifying promising treatments in specific subgroups of women with newly-diagnosed, high-risk, locally advanced breast cancer (Stage II/III). Quantum Leap, sponsor of the I-SPY 2 TRIAL, leads a pre-competitive consortium that includes the U.S. Food & Drug Administration (FDA), industry, patient advocates, philanthropic sponsors, and clinicians from 16 major U.S. cancer research centres.
The new I-SPY 2 treatment arm will evaluate SYD985 against standard of care therapy in Stage II/III early-stage, high-risk breast cancer patients, with a focus on tumours with heterogeneous and low HER2 expression. Byondis will supply the investigational drug and provide financial and regulatory support. Quantum Leap, as sponsor, will provide the clinical sites and clinical expertise.
SYD985 is Byondis’ most advanced ADC, targeting a range of HER2-positive cancers such as metastatic breast cancer (MBC) and endometrial cancer. The company is currently conducting a Phase III study of SYD985 (TULIP or SYD985.002) to compare its efficacy and safety to physician’s choice treatment in patients with HER2-positive unresectable locally advanced or metastatic breast cancer. Previously, the FDA granted fast track designation for SYD985 based on promising data from heavily pre-treated last-line HER2-positive MBC patients participating in a two-part Phase I clinical trial (SYD985.001).
SYD985 uses Byondis’ unique, proprietary linker-drug (LD) technology. Although marketed ADCs have improved therapeutic indices compared to classical non-targeted chemotherapeutic agents, there is still room for improvement.
SYD985 is comprised of the monoclonal antibody trastuzumab and a cleavable linker-drug called valine-citrulline-seco-DUocarmycin-hydroxyBenzamide-Azaindole (vc-seco-DUBA). The antibody part of SYD985 binds to HER2 on the surface of the cancer cell and the ADC is internalized by the cell. After proteolytic cleavage of the linker, the inactive cytotoxin is activated and DNA damage is induced, resulting in tumour cell death. SYD985 can be considered a form of targeted chemotherapy.
Dante Labs, Cambridge Cancer Genomics and Nonacus collaborate to provide precision oncology at scale
, /in Corona News, E-News /by 3wmediaDante Labs, a pioneer and leader in genomic testing, Cambridge Cancer Genomics (CCG.ai), a software developer specialising in data-driven precision oncology, and Nonacus, a provider of genetic testing products for precision medicine and liquid biopsy, have signed a collaboration agreement.
In a joint statement they said the partnership aims to build the most comprehensive and patient-centric tumour profiling service enabling improved cancer patient management, treatment and monitoring. By combining Dante Labs’ experience and capacity in delivering a sequencing service for both solid tumour and cell free circulating tumour DNA from liquid biopsies, Nonacus’ sensitive targeted pan-cancer NGS libraries, and CCG.ai’s industry leading AI powered software platform, OncOS, the companies will enable precision oncology at scale.
Improving outcomes for cancer patients means ensuring they have the right drug, at the right time to beat their cancer. This means understanding the molecular profile of the individual cancer and using that data to recommend treatments or clinical trials. Oncologists and clinical researchers will be able to send samples for processing to Dante Labs, who will use library preparation kits from Nonacus and software from CCG.ai to create a sample to report solution. If there are actionable mutations, the report will recommend the right treatments for those mutations, if there are novel or unactionable mutations, the software will also be able to match possible clinical trials.
Chris Sale, CEO of Nonacus, said: “Long turn-around time and lack of clinically oriented analysis are the main obstacles to fully deliver the potential of cancer genomics to patients. This partnership will provide the flexibility and accuracy that oncology professionals need to integrate cancer genomics into the care of their patients. The COVID pandemic has increased the backlog of genetic testing for cancer, potentially leaving many suspected cancers unconfirmed and treatments delayed. Dante Labs are one of the biggest clinical sequencing hub in Europe able to process large numbers of samples in high throughput. It is our hope that by combining AI software from CCG.ai and our library preparation kits, together we will be able to process samples and provide bioinformatic analysis critical to determining the best treatment path for patients.”
Start Codon accelerator showcases first cohort of start-up life science companies
, /in E-News /by 3wmediaStart Codon, a new model of life science and healthcare business accelerator, has announced its first cohort of start-up companies. Start Codon aims to minimise risk and translate early stage research into successful start-ups, ready for funding and partnership. Start Codon has worked closely with four life science and healthcare companies that were enrolled into the programme in February this year.
They are:
Start Codon plans to invest in and support up to 50 start-up companies over the next five years. The accelerator is now accepting applications for its second and third cohorts of companies. Early stage start-up companies in the life sciences and healthcare space are invited to apply via https://startcodon.co/application-form
ReactoMate DATUM — a user-friendly support system for laboratory scale reactions
, /in E-News /by 3wmediaThe ReactoMate DATUM from Asynt is a high quality, dual-rod stainless steel and aluminium support system built to ensure the stability and safety of your lab reactor. Sturdy, yet compact, the ReactoMate DATUM support system can accommodate a wide range of reaction vessels from 100 mL up to 5000 mL.
Designed with user-friendliness in mind, the ReactoMate DATUM support system incorporates a suite of innovative features.
Changing a vessel supported by the ReactoMate DATUM is as simple as “Clip & Click”. The novel neck clamp allows fast changeover between reactor vessel sizes thereby enabling simple reaction scale-up, whilst the ingenious mounting mechanism ensures excellent stability and alignment every time.
The Reactomate DATUM support system is fully compatible with all leading brands of overhead stirrers and circulator heating/cooling systems. Designed by chemists for chemists, low-friction polymer bearings line both the overhead stirrer alignment chuck and the neck support to ensure smooth and easy operation.
Ideally suited for use within a benchtop fume hood, adjustable feet allow you to level the ReactoMate DATUM support system ensuring stability and security while you work. Each DATUM system is also supplied with a moulded drip tray that fits perfectly within the base of the support, for safely catching any drips and spills from the reaction vessel during draining.
With a wide range of accessories and upgrades available, including drain manifolds and automation packages, the ReactoMate DATUM support system is the perfect all-rounder for laboratory scale reactions.
For more information, visit: www.asynt.com/product/reactomate-datum
Concarlo awarded patent for novel therapeutic peptide for drug-resistant breast cancer
, /in E-News /by 3wmediaConcarlo Holdings has received a US patent for IpY, a novel therapeutic peptide that addresses drug-resistant breast cancer by targeting a unique cellular pathway — p27Kip1. The patent is the latest step in Concarlo’s journey to commercialize revolutionary medicines for metastatic breast cancer.
Concarlo has also announced that a new provisional patent application has been filed for modified versions of the therapeutic peptide that are believed to exhibit enhanced bioavailability. Concarlo is a Brooklyn, New York-based biotechnology innovator dedicated to developing sophisticated, targeted therapies and diagnostics in the oncology space. The IpY technology is the first to address the high incidence of drug-refractory disease that develops with currently available CDK4 inhibitor (CDK4i) treatments. Such a solution has the potential to drastically increase overall survival of breast cancer patients.
The recent introduction of CDK4i drugs, a class of medicines that directly targets the CDK4/6 pathway implicated in many malignancies, has had a significant impact on the way in which metastatic breast cancer is managed. However, such therapeutics are associated with patients transitioning to a treatment-resistant form of the condition, despite initial extended periods of remission. Backed by more than 20 years of research and development expertise, Concarlo has developed IpY and a companion diagnostic, ApY, to effectively overcome the issue of CDK4i resistance and roll out a more targeted treatment approach for optimized patient outcomes.
“Despite the clinical efficacy of CDK4 inhibitors, we’re seeing that primary or secondary resistance to therapy is presenting a significant challenge to overall survival,” said Dr. Dominique Bridon, Chief Development Officer at Concarlo. “With the IpY technology and its unique mechanism of action, we’re effectively targeting CDK4 while simultaneously inhibiting another target — CDK2 — which has been found to be a key molecular player in the development of drug resistance. In doing so, we are the first company to successfully address the CDK4i resistance issue to provide long-term durable tumour arrest. Combined with its highly specific targeting and low toxicity profile, the positive impact of this drug on the breast cancer treatment landscape is hard to understate.”
Concarlo was formed in 2016 and is supported by a team of internationally renowned experts forming its Scientific Advisory Board. To date, the company has raised more than $3.1 million to support the development, improvement, and commercialization of its IpY and ApY technologies to bring a precision medicine approach to breast cancer management. The newly issued patent for IpY and the provisional patent application for modified versions of the peptide are the first key milestones in Concarlo’s plan to build an extensive patent estate to maintain market exclusivity for its clinically relevant therapeutics.
Sphere Fluidics, Heriot-Watt University collaborate to develop next generation droplet generator instrumentation
, /in E-News /by 3wmediaSphere Fluidics, a company developing single cell analysis systems underpinned by its patented picodroplet technology, and Heriot-Watt University, a specialist, pioneering Scottish University, have been awarded a Knowledge Transfer Partnership (KTP) grant from Innovate UK, the UK’s innovation agency. The grant will facilitate the development of novel droplet generator instrumentation, which will be used to expand Sphere Fluidics’ portfolio of microfluidic instruments for advanced biologics discovery and therapeutic cell line development.
Awarded to promote the collaboration of knowledge, technology and skills within the UK Knowledge Base, the KTP has been granted to Sphere Fluidics, in partnership with Dr. Graeme Whyte, Associate Professor at Heriot-Watt University. The two-year project will develop next-generation intelligent instrumentation and advance research across a range of picodroplet techniques, allowing scientists to discover rare cell phenotypes and to help to solve a range of biological questions ranging from antibody discovery to antimicrobial resistance, enzyme evolution and synthetic biology. The novel platform for semi-automated picodroplet production will be employed by the company to improve control of droplet production, using advanced imaging technology.
As part of the project, Dr. John McGrath has been appointed to Sphere Fluidics’ team as a Research Scientist in physics and engineering, to support the transfer of cutting-edge research into the company’s portfolio of single-cell analysis instruments, including for several new commercial products.
Dr. Marian Rehak, VP of Research and Development at Sphere Fluidics, said: “This innovative project with Heriot-Watt University, will bring together aspects of microfluidic and optical design, technology development and product design engineering to develop a new class of instrument for cell-based picodroplet discovery. We are delighted to have been awarded the KTP Fellowship and to welcome Dr. John McGrath to the Sphere Fluidics team. The work demonstrates the importance of collaboration between academic and industrial partners to support the advancement of novel microfluidic technologies for ground-breaking research.”
Dr. John McGrath, Research Scientist at Sphere Fluidics, commented: “I am thrilled to be working alongside commercial and academic leaders in the research and development of microfluidic instruments and technology. The ease of use and broader application set of the instrument to be developed in this project should lower the barrier to entry for a wide number of scientists, who are focused upon high-throughput screening, synthetic biology, gene editing, and antimicrobial resistance workflows. The technology has the potential to be a key driver in increasing the uptake of picodroplet microfluidic instruments.”