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Assays and Performance

Point-of-Care immunoassays with unmatched sensitivity

Cardiac Troponin I 

Novilux's Cardiac Troponin I (cTnI) assay is poised for commercialization. Key elements of the cTnI assay have been tested and de-risked for eventual clinical trial and FDA submission.  The SMC assay has been run on 100,000's of patient samples in CLIA format. Thousands of blanked, endogenous and spiked runs on the Phoenix platform have been run to assess POC performance. When run on the Phoenix systems the assay is an order of magnitude more sensitive than the latest generation of high-sensitivity instruments used in the hospital's Centralized Reference Laboratory, and 100X more sensitive than other POC systems. Results are available quickly and can be optimized to meet the specific needs of the acute setting.

Measurement Range  0.2 ng/L to 15,000 ng/L

Measured concentration (ng/L)

cTnI SinglePlex Graphs.png

All samples

Lower  3  samples

Expected concentration (ng/L)

Analytical Sensitivity

Repeated studies with whole blood and plasma demonstrate ultra-high sensitivity. The graphic to the left shows a dose response for cTnI serially diluted in EDTA anti-coagulated plasma and measured on multiple Novilux Phoenix point-of-care instruments. The upper plot shows the full range of cTnI samples between 0 ng/L (cTnI depleted blanks) and 15,000 ng/L. The lower plot is scaled to show the lower three concentrations. The table below provides the numerical results for the same study and lists LoBDQ values calculated in accordance with CLSI EP17-A protocols. 

cTnI SinglePlex Data Table.png

All samples input, processed and analyzed with a 12 minute Time to Result

Multiplexed Cardiac Panel
(cTnI, NT-proBNP, D-dimer, CK-MB)

Novilux SMC™ technology can multiplex assays both spatially and optically. Multiplexed panels are used to quickly rule-in or rule-out various conditions at the molecular level that may present with the same symptoms at the anatomical level. For example, Acute Coronary Syndrome (ACS), Heart Failure (HF), and Pulmonary Embolism (PE) all share shortness-of-breath as a symptom at the time of presentation to the hospital Emergency Department. cTnI and CK-MB are often used to aid in the diagnosis of ACS (myocardial infarction in particular), NT-proBNP aids in the diagnosis HF and D-Dimer is used to help diagnose PE. Providing all four markers simultaneously can be of enormous benefit to differentiate diagnosis of cardiac conditions when overlapping symptoms are present.

NT-proBNP Measurement Range  2.6 ng/L to 105,000 ng/L

Duplex NT-proBNP.png

cTnI Measurement Range  0.2 ng/L to 15,000 ng/L

Duplex cTnI.png

D-dimer Measurement Range  4.4 ng/mL to 5,000 ng/mL

D-dimer 01.png

CK-MB Measurement Range  0.04 ng/mL to 150 ng/mL

CK-MB 01.png

Multiplexed Cardiac Panel
cTnI   /   NT-proBNP  -  Phoenix 

The graphs on the left show assay results for a 3-channel, optically multiplexed, cardiac panel including Cardiac Troponin I (cTnI), NT-proBNP, and a positive control run on Phoenix POC system. The same cTnI assay described above is tested in the RED channel, with an early-stage development assay for NT-proBNP tested in the GREEN channel. Both cTnI and NT-proBNP have been serially diluted ten times from concentrations of 15,000 ng/L and 105,000 ng/L respectively. The positive control, assessed in the NIR channel, was set at a constant level in all 125 samples tested and generated a CV of 2.6%.

 

The inset in each graph in the lower right-hand corner shows the lowest three samples including cTnI / NT-proBNP depleted blanks, endogenous samples and the next lowest spiked sample in the dilution series.

 

The analytical sensitivities for cTnI and NT-proBNP  are shown in the table below:

Triplex cTnI and NT-proBNP.png

Notes: 

  • Results were obtained by pooling data from 125 samples across three Phoenix POC systems operating with a ~12 minute Time-to-Result. 

  • Mean concentration of Endogenous cTnI at 1.2 ng/L is > 15 SDEV above cTnI depleted blank.

  • Mean concentration of Endogenous NT-proBNP at 29 ng/L is > 60 SDEV above NT-proBNP depleted blank. 

D-dimer  /   CK-MB    -   RDP

The graphs to the left show results for D-dimer and CK-MB run in the GREEN and RED channels. These assays are still in early development and were run on the Novilux RDP platform. Unlike the Phoenix POC systems the RDP can run assays with reagents in liquid format in a highly parallel processing mode. When performance metrics have been met, the reagents are converted to lyophilized format for porting to the Phoenix POC systems.

 

The RDP enables rapid development of assays running 1000’s of samples and making needed adjustments to the assay. This is often difficult on POC platforms since the underlying technology does not lend itself to highly parallel processing. This is not the case for Novilux SMC™ technology where the same technology can be used in Point-of-Care systems, the research space and in highly parallelized diagnostics Reference Laboratory Platforms.

 

As shown in the graphs to the left and in the table below, the D-dimer and CK-MB assays both demonstrate sensitivities and measurement ranges that are superior to existing Reference Laboratory instruments. Both of these assays are close to being lyophilized and ported to the Phoenix POC platform.  

RDP D-dimer and CK-MB.png

Notes: 

  • The complete cardiac panel is a 4-plex assay spatially and optically multiplexed including cTnI, NT-proBNP, D-dimer and CK-MB.

  • D-dimer and CK-MB assays are currently in development on the RDP platform and will be ported to the Phoenix POC instruments. 

  • When complete all four assays will run simultaneously on the Phoenix instruments with cTnI and NT-proBNP processing in one circuit while D-dimer and CK-MB are being processed simultaneously in a second circuit. 

SARS-CoV-2 Antigen   (demonstration)

Novilux SMC™ technology is well suited to just about any immunoassay including the SARS-CoV-2 antigen. The SARS-CoV-2 assay is complementary to the cTnI assay owing to the established connection between the severity of COVID-19 infection and cardiomyocyte damage. Most SARS-CoV-2 antigen tests used today are lateral flow immunoassays with Emergency Use Authorization (EUA) by the US FDA. The SMC™ SARS-CoV-2 assay developed at Novilux for demonstration purposes is 100-fold more sensitive than these tests.

SARS-CoV-2 Antigen LoD

The graph on the right shows the relative sensitivity of a Novilux early prototype of the SARS-CoV-2 antigen SMC assay (in blue at 0.6 pg/ml) compared to many of the commercially available tests. These assays have a median LoD of 100pg/ml and with only one exception all have LoD's higher than 10 pg/mL. Other than Novilux, only the Quanterix Simoa has an LoD of less than 1 pg/ml. However, the Simoa assay cannot be done at Point-of-Care, and results take more than an hour after sending to a Centralized Lab running an LDT.

Note:  These results are for demonstration purposes. The results were generated using the Novilux RDP assay development platform. The assay has not been ported to the Phoenix Point-of-Care system.

SARS-CoV-2 Ag LoDs 4.PNG

Growing Ecosytem for Biomarker Development

One of Novilux out-licenses for the SMC™ technology is to MilliporeSigma (Merck KGaA) for the Research Use Only market. That license has enabled a large and growing installed base of SMCxPRO™ instruments, MilliporeSigma assay kits, and 1000's of researchers in the field developing new biomarkers based on SMC™ discoveries.

As groundbreaking new clinical research validates important new biomarkers using SMC™ technology, Novilux is poised to collaborate and implement those biomarkers on its Phoenix Point-of-Care platform.

> 130 SMC Assays to Date

The graphic to the right shows a collection of 54 of the more than 130 ultrasensitive immunoassays developed to date on SMC™ platforms. The biomarkers enumerated here are important for monitoring a broad array of health conditions, many of which could be developed into diagnostic tests or panels.

   

An ecosystem for ultrasensitive assays is now taking shape where commercial Life Science discoveries can become validated diagnostic tests/panels via porting to Novilux ultra-high sensitivity, low-cost diagnostic platforms.

Infographic Footnote Key. Assays developed or sold by:

1. Novilux

2. Singulex

3. MilliporeSigma

4. External, SMCxPRO user developed 

Assays Infographic v4.png
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