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Core Laboratory for Clinical Studies (CLCS) is Washington University for supporting research across all departments.
The Core is developing new assays with the Singulex Erenna Reader. Only a few similar instruments currently are installed in academic labs. The Singulex system is based on digital single molecule counting. It can be thought of as a “smart” ELISA reader with a greatly extended lower analytical measurement range. The purpose of this research is to increase sensitivity of the assay and to improve reproducibility at the limits of measurement.
SAN FRANCISCO — Monitoring levels of cardiac troponin I identified post-ACS patients at heightened risk for future death from CVD or HF, according to new data from the PROVE-IT TIMI 22 trial.
"Using a next-generation investigational assay for cardiac troponin, we found that the vast majority of patients who were stable after ACS 4 months previously had detectable levels of circulating troponin and that that level was strongly associated with subsequent cardiac events," David A. Morrow, MD, director of the Levine Cardiac Intensive Care Unit at Brigham and Women's Hospital, told Cardiology Today.
SAN FRANCISCO—Serial measurements of cardiac troponin using an investigational high-sensitivity assay enhanced risk stratification for cardiovascular disease and heart failure in patients with stable ischemic heart disease, according to an analysis of the PROVE IT-TIMI 22 study, presented March 9 at the American College of Cardiology’s (ACC) scientific session.
Cardiac troponin is an emerging prognostic biomarker in patients with stable ischemic heart disease. In this study, lead investigator Ryan G. O'Malley, MD, of Brigham and Women's Hospital in Boston, and colleagues assessed the prognostic performance of serial measurements of an investigational high-sensitivity cardiac troponin assay in patients with stable ischemic heart disease.
Serial measurements of cTnI using an investigational high-sensitivity assay enhanced risk stratification for CVD/HF in pts with SIHD.
Singulex Inc, Alameda, Calif, reports that new data from the Thrombolysis in Myocardial Infarction (TIMI) Study Group to be presented at the upcoming American College of Cardiology Annual Scientific Session & Expo have shown that monitoring precise blood levels of the biomarker cardiac troponin I (cTnI) is helpful in identifying post-acute coronary syndrome patients who are at increased risk for future death from cardiovascular disease or heart failure.
The Singulex high-precision cardiac troponin I digital research-use only assay, which enabled the measurement of cTnI at levels below 10 pg/mL, was used for the study that will be presented at the event.
"The study shows that serial monitoring cTnI levels over time using a high-precision assay provided additional prognostic information, enabling us to assess future risk in post-acute patients," says David A. Morrow, MD, study investigator and director, TIMI Biomarker Program and director, Levine Cardiac Intensive Care Unit, Brigham and Women's Hospital and associate professor, Harvard Medical School.
"Patients with elevated cTnI or whose cTnI increased in the course of 90 days were at greater risk for cardiovascular death and heart failure," Morrow adds. Previous data from the PROVE IT-TIMI 22 trial have shown that these patients with elevated cTnI appear to have the most to gain from intensive versus moderate-dose statin therapy.
Singulex has collaborated with Morrow and his group at Brigham and Women's Hospital for several years.
The results could have implications for the future adoption by cardiologists of high-sensitivity troponin assays currently being developed by biotech firms like Singulex and Nanosphere.
Alameda, Calif.-based biotech firm Singulex this week announced it has been granted a US patent related to a cardiovascular disease biomarker assay using its Erenna single-molecule immunoassay platform.
The patent follows on one the company received last year for the Erenna platform itself, and comes as it begins efforts to develop the assay as a clinical diagnostic that it hopes to submit for US Food and Drug Administration approval within the next two years.
The Erenna platform uses a capillary flow system to enable counting of single molecules in immunoassays. According to CEO Philippe Goix, the technology can quantitate target proteins with sensitivity one to two orders of magnitude greater than a standard ELISA. That, he noted, allows researchers to quantify known protein biomarkers at previously undetectable levels – potentially opening up new uses for established markers.
US Patent No. 7,572,640, awarded in August 2009, covers the flow system method used in the Erenna platform to achieve single-molecule counting. Singulex's second patent, US No. 7,838,250, awarded last month, covers the measurement of the protein cardiac troponin-I at low-abundance concentrations – the key to the cardiovascular disease diagnostic the company is developing.
Troponin-I is a well known biomarker for cardiac tissue damage, often used in acute settings to determine whether a patient has had a heart attack. It has been less useful for long-term monitoring purposes, however, because of the difficulty in measuring it at low enough concentrations to establish a baseline for normal subjects.
This, Goix told ProteoMonitor, is where Singulex hopes to apply the Erenna technology.
"If you can pick up troponin-I [in a patient], you can rule them in [as having suffered heart damage], but it's very difficult to rule anyone out, because to rule them out you have to measure [troponin-I] at normal levels," he said. "With our technology we can measure the concentration of [troponin-I] at very low levels, and now we can use that information to really monitor people and the risk that they will develop cardiovascular disease."
Conventional ELISAs can quantify troponin-I at concentrations around 30 to 40 picograms per mL, Goix said. The Erenna platform, on the other hand, measures the protein in a range of approximately 1 to 10 picograms per mL.
Equally important for cardiovascular monitoring purposes, he said, is the platform's resolution.
"Say a person measures [troponin-I levels] of 2 picograms. Normally it stays at that level give or take 50 percent. Now let's say you measure it at 5 picograms. That's 250 percent higher – so that's big," he said. "We have a technology that can measure not only 2 picograms, but that can measure [changes from] 2 picograms to 5 picograms to 10 picograms very accurately, and it turns out that there is a lot of clinical utility to that."
Singulex is not alone in the cardiovascular biomarker space. This week a study published in the European Journal of Heart Failure demonstrated that the Verisens troponin-I assay from molecular diagnostics firm Nanosphere was able to stratify patients at risk of heart failure. Other companies like BG Medicine (PM 12/03/2010) and Insilicos (PM 09/24/2010) are also investigating biomarkers for cardiovascular disease.
As part of its planned FDA submission, Singulex is working on a new version of the Erenna platform for use in clinical settings, including, potentially, at the point of care. Currently, the company offers the troponin-I assay out of its CLIA-certified lab. It offers CLIA-certified assays for the proteins IL-17A, IL-6, and TNFα, as well, which are related to inflammation and cytokine pathways. Additionally it sells a variety of non-CLIA assays for use on the Erenna platform that Goix characterized as "research grade."
Singulex also works with a variety of large pharmaceutical companies, he said, including Roche, Novartis, and Pfizer. In these partnerships, the company applies its technology to the high-sensitivity quantification of biomarkers that these firms have developed as part of their drug development efforts.
"We're very complementary to, say, a mass spec company where you have, in discovery, say, 500 biomarkers you need to look at," he said. "The next level is where you have maybe 10 biomarkers and you want maybe two fully validated. You'd use our technology for that. We develop a clinical-grade answer; we're more in the diagnostic-grade development."
Bringing high-sensitivity quantification to established markers has been a trend for single-molecule detection firms. Much like Singulex's work with troponin-I, single-molecule analysis company Quanterix has used its Single Molecule Array – or SiMoA – technology to measure the established biomarker prostate specific antigen in the blood of radical prostatectomy patients at concentrations far lower than that allowed by conventional ELISA-based tests. A study using the company's technology published in the June issue of Nature Biotechnology suggested that such high-sensitivity measurements of PSA could be useful in assessing the risk of cancer recurrence (PM 06/11/2010).
In addition to cardiovascular disease, Singulex is now looking to apply its platform to autoimmune disorders, Goix said. The company is working with researchers at New York University on rheumatoid arthritis, and in September won part of a €2.2 million ($2.9 million) grant from France's Single Inter-Ministry Fund for an R&D program headed by Indicia Biotechnology in collaboration with the Hospital of Lyon, TxCell, and Novartis, to develop biomarkers for Crohn's disease on the Erenna system. It began work on the project this month.
The company is also working to develop offerings in oncology, Goix said, as well as considering assays aimed at Alzheimer's disease. In particular, the Erenna platform has had success detecting the Alzheimer's cerebrospinal fluid biomarker amyloid-beta 42 in serum, raising the possibility of a blood-based test that could obviate the need for more invasive lumbar punctures.
"We have plans to be a diagnostic player. We definitely have plans to go that route," he said. He noted, however, that right now the company is focused on building a profitable business based on its assay offerings and services work for academic and pharma partners.
Singulex is venture funded, with backing from firms JAFCO, OrbiMed Advisors, and Fisk Ventures. It closed a $19 million financing round in January 2009. Goix said it would likely raise additional funds as it worked toward commercializing the patented technologies but declined to give further details.
We introduce here our highly sensitive immunoassay for troponin I. We are beginning to see the next generation of assays that can report measurable troponin concentrations in healthy subjects. Our Erenna® immunoassay system measures as little as 1.7 ng/L of troponin I with 10% CV, and the 99th percentile in a group of apparently healthy subjects was 7 ng/L.
A single molecule counting approach to early disease detection developed by Singulex targets low abundance proteins in the bloodstream and can detect protein biomarkers at pg/mL concentrations. The company is evaluating the utility of cardiac troponin-I as a biomarker for acute myocardial infarction, and of plasma cytokines in inflammation.
Already playing first chair among tests used to detect acute myocardial infarction, cardiac troponin may have other instrumental roles in its future. New, more sensitive troponin assays on the near horizon hold the promise of improving diagnosis in acute coronary syndrome, as well as risk stratifying patients in numerous settings, including potentially primary care.
Technological advances in life sciences can feed the early-stage assay development pipeline and can be transitioned into the clinical diagnostics setting.
Proteins are the machinery that drive most biological processes: however, measuring their concentrations at low levels has been historically impeded by a lack of appropriately sensitive technology. Advances in immunoassay technology, such as single molecule counting, have provided orders of magnitude of increased sensitivity. These novel technologies are enabling new discoveries and a clearer understanding of previously intractable biology. Here we discuss how single molecule counting technology can brighten the future of translational research, drug discovery and personalised medicine.
If one company's disaster can transform an industry, the withdrawal of Merck's $2.5 billion painkiller Vioxx in 2004 may become the textbook example.
Primarily, the removal of Vioxx from the market due to heart risks spurred a sweeping examination of the pharmaceutical industry and its regulation. But it also enriched the soil for a new generation of companies trying to update technology that detects whether new drugs may be harmful to the heart....
...Some companies also improve existing assays. Singulex, Hayward, Calif., for example, developed a technique for single-molecule detection that “extends the dynamic range of immunoassays by one to two orders of magnitude,” says Philippe Goix, PhD, chief executive officer. “If you have an immunoassay, we can quickly integrate it into our platform without drastically changing it.” Singulex is already developing ways to use this technology in clinical trials. For one thing, its high sensitivity could measure normal levels of hard-to-measure proteins, and then look for changes after administering a drug during a clinical trial....
In 2004, the U.S. Food and Drug Administration released a white paper, now known as the Critical Path Initiative, which challenged the pharmaceutical industry to reduce the time (12Š15 years) and expense (approximately $1Š2 billion) required to bring a drug to market. Biomarkers were seen as key to meeting this challenge with a promise of providing more effective drug and target identification, safety and toxicity monitoring, and patient identification and stratification. With unprecedented sensitivity and dynamic range, the technology described in this article helps fulfill the promise of biomarkers through its advances in single molecule detection.
A tremendous amount of progress has been made in developing assays. Most of these advances have been in genomic level disease-screening assays that provide yes/no answers. Because one gene can trigger the formation of many different proteins, protein biomarkers offer a wealth of valuable information for clinical development and monitoring. The challenge is to produce robust proteomic assays that are also clinically validated. In this article, IVD editor Richard Park and Singulex president Philippe Goix discuss:
OVERVIEW: Early identification of adverse heart events is invaluableŃnot only in clinical diagnostic settings, but in clinical trials as well. The Singulex Erenna Immunoassay System detects small changes in concentration levels of cTnI well below the current and standard detectable limit of all other Troponin assays. In fact, the assay is able to measure the level of troponin I in normal individualsŃsomething Singulex president Philippe Goix says has never been possible before. With the cTnI assay and other similarly sensitive assays, Singulex looks to improve clinic diagnostics as well as helping pharmaceutical companies prevent late-stage drug development failures.
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