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HomeSMC® High Sensitivity Immunoassay KitsAn Overview of SMC® Ultrasensitive Immunoassay Technology

An Overview of SMC® Ultrasensitive Immunoassay Technology

How can ultrasensitive immunoassay technology advance your studies? In this article, learn how SMC® technology can be used in research areas including immunology, cardiotoxicity, and neuroscience to provide more comprehensive biological insights by revealing signals that fall below the detection limits of traditional immunoassays.

Section Overview

What is SMC® Technology?

SMC® technology is an advanced ultrasensitive immunoassay technology that gives researchers the power to detect biomarkers that were previously undetectable down to femtogram/mL levels. It provides an indispensable tool in the researcher’s arsenal to help move novel biology forward, fueling the discovery and development of new therapeutics. SMC® technology uses a basic sandwich immunoassay format utilizing two antibodies specific to the analyte of interest: a capture antibody coated on a magnetic bead or plate and a detection antibody conjugated to a fluorescent protein. Following sandwich formation, a proprietary elution step dissociates the immunocomplex, allowing for detection of free-floating, labeled detection antibodies. This enables the high sensitivity measurement of larger molecules and compatibility with variable assay formats. Single or dual analyte analyses can be performed with the FemtoQuest® system. Only SMC® immunoassay technology allows researchers to use both bead-based and plate-based methods, thus offering unmatched flexibility in assay design.

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SMC® Technology Workflow

Traditional immunoassays have limitations such as matrix effects, dynamic range, and sensitivity that can impact the ability to accurately measure or even detect low abundant proteins. These limitations can lead to false positives or negatives, incorrect assumptions, and missed opportunities, overlooking critical early disease biomarkers. The differentiator of the patented SMC® technology from other high sensitivity platforms that read signal from a surface-bound complex, is its unique elution step which breaks apart the immunoassay complex, allowing for counting of individual molecules. The eluate is then transferred to a SMC® 384-well read plate, to further isolate and concentrate detection antibodies for quantification, improving signal uniformity. Not only does this remove other assay components, like beads and unbound antibodies, that can contribute to high background signal, but it makes it one of the only ultrasensitive immunoassay platforms architecturally immune to steric hindrance from large proteins, biologics, and drugs, making it an ideal choice for immunogenicity studies.

The SMC® technology workflow is described below. In summary, analytes of interest bind to a capture antibody conjugated to magnetic beads. Then, a fluorescently labeled detection antibody forms a sandwich complex with the analyte and capture beads. Following complex formation, a proprietary elution step is used to chemically disassociate individual detection antibodies and is transferred to the SMC® 384-well read plate for reliable, high sensitivity quantification. The plate is loaded into the FemtoQuest® instrument where a laser excites the fluorescent-labeled detection antibody as it passes through a narrow interrogation window. Individual photons are captured by an avalanche photodiode and the signal is recorded. This allows for the digital quantification of individual molecules. Analyte concentrations in the unknown samples are calculated using the corresponding standard curve.

The steps in the SMC® Technology Workflow include:

Graphic showing yellow, magenta, and blue ovals representing analyte 1, analyte 2, and other proteins in a sample.

Sample Prep

Analyte(s) of interest in solution.

    Illustration showing a yellow oval representing analyte 1 and a pink oval representing analyte 2, each bound to antibodies coated to a surface.

    Capture analyte(s) on plate or bead.

      Illustration showing a yellow oval representing analyte 1 and a pink oval representing analyte 2, each 'sandwiched' between an antibody coated to a surface and a fluorescent-labeled antibody.

      Antibodies translate each biomarker into a signal.

        Illustration showing disassociation of antibody/biomarker/fluorescent-labeled antibody complexes.

        Complex is chemically broken apart.

          Illustration depicting two fluorescent-labeled antibody/analyte complexes within a beige circle representing the interrogation window.

          Sample is detected by laser and detection tags are counted.

            The unique features of the SMC® immunoassay, including available custom assay development and off-the-shelf kits, result in reliable quantification of proteins in low abundance. With this technology, researchers can:

            • Easily self-develop assays with flexible customization options to measure novel, elusive biomarkers
            • Retain samples and re-read plates, since signal acquisition occurs from the bottom of the read plate
            • Rely on robust antibody binding, with no spatial constraints
            • Detect both large and small molecules easily with a broad dynamic range
            • Access a large portfolio of available, verified, lot-matched kits
            • Reduce program costs and improve efficiency with the simple-to-use FemtoQuest® instrument
            • Reduce manual hands-on time by automating SMC® assays with ready-to-go protocols on the plug-and-play AAW™ workstation 

            Ultrasensitive Biomarker Detection Platform

            SMC® technology offers an ultrasensitive biomarker detection platform with the flexible FemtoQuest® system. Table 1 compares running assays on this platform to running traditional ELISAs.

            Ancillary Equipment and Kits to Enhance the SMC® Workflow

            To enhance the FemtoQuest® platform, we also offer ancillary equipment and kits specifically for SMC® technology, such as:

            Research Applications

            Ultrasensitive immunoassays can enhance research in a variety of fields because the improved sensitivity allows researchers to dive deeper into their studies. Research applications of SMC® technology include the areas of:

            Frequently Cited Advantages of SMC® Technology

            SMC® technology provides maximum immunoassay performance while following a workflow very similar to traditional ELISA technology.

            Traditional ELISA methodologies demonstrate limitations in sensitivity and dynamic range, typically require high sample volumes, and are susceptible to matrix effects. Combined, these factors reduce the utility of traditional ELISAs for the detection of low-abundant proteins and endogenous biomarker levels in healthy subjects, thus hampering statistical analysis among study groups. By adapting an ELISA workflow, SMC® technology achieves improved signal-to-noise ratios over traditional immunoassay technologies, thus providing quantification at both low and high levels of expression in one complete system. Digital counting of fluorescent events improves the assay sensitivity and extends the assay dynamic range beyond what can be achieved with traditional immunoassays.

            Because SMC® immunoassay technology can reach fg/mL sensitivity ranges, this platform offers the ability to dilute pre-clinical samples, when only low sample volumes are available. 

            Users are fully supported by Specialists, as well as dedicated technical support teams.

            We understand the SMC® platform is an important investment for research labs and we are committed to ensuring the success of its users. Regardless of the types of assays being used, all SMC® users are fully supported by Specialists who have experience working with researchers from a broad range of lab types, including academic, government, biotech, pharma, CRO, and regulated labs.

            The SMC® Assay Development Kit and the FemtoQuest® system make assay development simple.

            Don’t see your biomarker of interest in our expanding portfolio of SMC® high sensitivity kits? We have an easy solution for that. Customize your way with our SMC® Bead-Based Assay Development Kit (Prod. No. 03-0222-00). This kit provides an all-in-one solution to streamline your assay development process. It contains all assay reagents and clearly defined protocols to create application-specific immunoassays tailored to your research needs. Use this kit in conjunction with the SMC® Assay Buffer Optimization Kit (Prod. No. 03-0221-00) to further improve custom assay performance. Robustly and affordably detect your low-abundant biomarkers using our open platform FemtoQuest® system with SMC® technology.

            The SMC® platform is a versatile system that can be used in multiple study types, including biomarker assessment, bioanalytical work such as pharmacokinetic and pharmacodynamic studies, and immunogenicity testing.

            Complimenting a menu of off-the-shelf, verified assay kits, our Custom Assay Development and Innovation (CADI) team can be contracted to perform custom assay development and sample testing services at their site in St. Louis, MO, USA. Request more information about our custom services on our CADI page.

            SMC® assays are available in bead-based formats and can be developed in both plate-based and bead-based formats by our CADI team.

            The proprietary SMC® technology allows scientists to measure proteins with increased precision, enabling unparalleled quantification at low and high abundant levels of expression. The flexible SMC® immunoassay system acquires data from both plate-based assays and bead-based assays, providing a choice of format depending on budget and quantification requirements. Learn more about how we develop custom SMC® assays in our article on CADI services.

            The SMC® 384-well read plate is pre-qualified and designed for optimal performance. It delivers reproducible results and supports consistent data generation across studies. This high-throughput platform allows researchers to perform an entire SMC® assay from sample prep through data analysis in one day.

            FemtoQuest® system can be integrated with the AAW™ Automated Assay Workstation to reduce hands-on time and ensure robustness and reproducibility.

            Automation of SMC® immunoassays is desirable so that researchers can focus on other high-value activities and increase overall efficiency. The AAW™ automated assay workstation is a modular robotic liquid handler powered by Opentrons®, that is designed for high-throughput and technical workflows such as SMC® high sensitivity immunoassays. Automating SMC® immunoassays with the AAW™ automated assay workstation helps customers achieve key performance metrics while reducing manual intervention steps.

            Integrated analysis with Belysa® curve fitting software.

            The FemtoQuest® system uses in-house developed software with optimized scan time and algorithms to improve variability, limit of detection and dynamic range for enhanced sample discrimination. The integration with the Belysa® Curve Fitting Software provides a user-friendly experience for data analysis.

            For labs operating in a regulated environment, the FemtoQuest® system generates a signal data stream that can be imported into Laboratory Management Systems, such as WATSON.

            Tips and Tricks for Running SMC® Assays

            Have questions about running SMC® assays? Check out helpful tips and tricks in our guide to high sensitivity biomarker analysis with SMC® technology.

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            Publications Using SMC® Technology

            See how researchers are using SMC® technology in the publications list below organized by research area.

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            For Research Use Only. Not For Use In Diagnostic Procedures.

            Curious for more? Learn about the capabilities of our multiplex immunoassays on our MILLIPLEX® multiplex assay overview page.

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            To see the most updated list of SMC® kits, visit SMC® High Sensitivity Immunoassay Kits.

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