Next Generation Immunodiagnostics is Driven by Innovative Single Molecule Counting Technology
Technology & Science Jimmy Chen
Singulex’s SMC technology has been validated in more than 110 peer-reviewed published studies involving over 100,000 patients. These peer-reviewed publications, ongoing research applications, and growing clinical adoption validate the scientific integrity of SMC technology.
Single Molecule Counting technology is 100 times more sensitive than contemporary immunoassay platforms, enabling unprecedented high precision and digital detection of low abundance biomarkers. This sensitivity reveals what couldn’t be detected before: the power to distinguish people who have disease from those who do not, before symptoms are apparent, leading to improved outcomes 1, 2
SMC™ technology can be applied to existing and newly identified biomarkers to aid physician decision making in cardiovascular, oncology, infectious disease, inflammation/immunology, and CNS applications.
Singulex technology couples proprietary digital SMC technology with robust microparticle-based immunoassays to provide higher sensitivity and broader dynamic range than contemporary immunoassay systems 3. A biological sample (e.g. blood plasma) containing the analyte of interest is combined with paramagnetic micro-particles coated with capture antibodies and detection antibodies labeled with fluorescent dye. After a brief incubation, the micro-particles are washed to remove any excess detection antibody and then subjected to conditions that disrupt the sandwich complex formed on the micro-particles. The resulting eluate, which contains the fluorescently-labeled detection antibodies, is separated from the micro-particles and analyzed using SMC technology. Single fluorescently-labeled molecules that pass through a small interrogation space created by a confocal detection system (see below) generate intense flashes of light as they are illuminated by a laser.
The image to the right depicts the confocal microscopic optics utilized in SMC™ technology. The optics rotate and scan through the bottom of a detection vessel. The intensity of fluorescence encountered in the interrogation space (orange dot) is captured as a function of time. The measured fluorescence is sequestered in 100 micro second time bins as shown below. A threshold of signal intensity (dotted line) is applied across all time bins and only bins with fluorescence above the threshold are counted as detected digital events. Each of these counted digital events represents a single fluorescently-labeled molecule. The sum of digital events is equivalent to the concentration of analyte in the original biological sample. The SMC technology provides a reading range of more than 6 Logs.
Proprietary Intellectual Property
Singulex’s pioneering research has yielded a broad and deep global intellectual property portfolio that includes both SMC technology as well as numerous biomarkers for cardiovascular, oncology, infectious disease, and inflammation/immunology applications.
1 Apple F, et al. Clin Chem. 2012; 58: 930-935. 2 Neumann J, et al. PLOS one. 2014; 9: 3: e90063 3 Todd J, et al. Clin Chem.2007; 53: 1990-1995