Trace® is an optical electromechanical biomarker detector with a measuring range between pg/mL to mg/mL

trace® measures fluorescence and/or absorbance changes in a reaction chemistry mix. One drop of blood is added to the cartridge, which is subsequently placed into the device. Each cartridge contains a radio-frequency-identifier (RFID) chip that transfers assay related information to the device, which is now ready to run the test.

trace® filtrates plasma from whole blood and via a microfluidics system transfers the plasma further into chemistry containers that are separately injectable into a cuvette, where the final reaction and measurement takes place in a temperature controlled environment.

Trace® - fluorescence and absorbance detection. The figure below illustrates the reaction principle.

Test Procedure (universal)

  1. Add blood to inlet
  2. Close cartridge lid
  3. Insert cartridge into instrument and press "start"
    1. Close drawer
    2. Assay protocol is identified by RFID chip
    3. Assay execution starts automatically
  4. Wait for result
    1. Results are shown on display, smart device or computer (Bluetooth)
    2. Results are stored in memory
    3. Optional: Results are transferred to central database (WiFi or GSM)

Trace® General Biomarker Assay Principle

Trace® Lipids

Trace® measures Total Cholesterol (CHOL), High Density Lipoprotein (HDL), non-HDL, Triglycerides (TG) and calculate values for Low Density Lipoprotein (LDL). Following the blood-to-plasma filtration, enzymes are serially injected into the reaction cuvette at different time points and the CHOL, HDL, non-HDL and TG are measured by a stepwise enzymatic end-point protocol that generates absorbance changes that correlates to concentration.

The figure below illustrates the methodology for the Trace® Lipids

The lipids protocol execute the following 5 primary steps:

Blocking of non-HDL
Plasma transferred to proprietary R1 solution in the main cuvette.

HDL assay
R2 proprietary solution added and device measures the absorbance difference between R1-R2 which correlates to the HDL concentration.

TC assay
R3 proprietary solution is added and device measure the absorbance difference between R2-R3 which correlates to the non HDL concentration.

TG assay
R4 proprietary solution is added and device measures the absorbance difference between R3-R4 which correlates to the TG concentration.

LDL calculation
Friedewald formula: LDL = CHOL- HDL-TG/5 (mg/dL).

Lipids cartridge actions

Trace® HbA1c

trace® transfers a precise volume of whole blood into the reaction cuvette. A dry fluorescent borate conjugate (FBC) is injected into the reaction cuvette and mixed with a proprietary enhancement and lysis buffer. A decrease in fluorescence signal is obtained through a specific binding of the borate fluorescence molecule to the glycated portion of the haemoglobin molecules. The decrease in the fluorescence channel correlates to the HbA1c concentration. Post theHbA1c measurement total haemoglobin is measured by injecting sodium lauryl sulphate (SLS) into the cuvette where it is mixed with the solution. The SLS oxidizes the total haemoglobin (tHb) and forms the complex sodium lauryl sulphate-tHb, the colour of this complex is then measured in absorbance channel. The colour complex is proportional to the total haemoglobin concentration in the sample. HbA1c is then calculated as follows: HbA1c = HbA1c(s) / tHband presented in IFCC a unit (mmoI/moI). (IFCC - international Federation of Clinical Chemistry).

The figure below illustrates the protocol executed by Trace® HbA1c

HbA1c cartridge actions

Trace® Companion Diagnostics Assay Principle

Trace® — a universal TNFα inhibitor assay

Whole blood is entered from fingertip into blood inlet. Following the blood-to-plasma filtration trace® uses an absorbance based particle-enchanced immuno turbidimetric method for measuring the biological TNFα inhibitor drug molecules. Latex particles coated with the TNFα molecule are released into the reaction cuvette where the TNFα inhibtor drug is present, an agglutination reaction then initates. The reaction causes absorbance changes that are proportional to the drug concentration. Each drug has its own calibration factor related to each particular drug’s affinity towards the TNFα ligand molecule.

trace® - a universal TNFα inhibitor assay
The Figure below illustrates the trace® CDx principle

Trace® CDx ADA Assay
-anti-drug-antibodies (ADA) towards TNFα inhibitor drug

Having measured the drugconcentration with Trace® CDx Biological Drug Assay a second cartridge is applied — Trace® CDx ADA Assay — this cartridge contains identical assay reagents plus an additional spiked concentration of the particular biological drug tested for neutralizing anti-drug-antibodies. If the CDx ADA Assay gives significant less concentration of the initial drug assay measured it is likely due to the presence of neutralizing anti-drug-antibodies.

Example of how Trace® CDx ADA Assay principle works:

Cartridge 1: Apply trace® CDx Biological Drug Assay and measure drug concentration. Obtained result e.g. 2μg/mL

Cartridge 2: Apply trace® CDx ADA Assay to determine ADA presence (for the particular drug the cartridge contains 3μg/mL of the drug).

Results:
= 5 μg/mL = no neutralizing anti-drug-antibodies.
< 5μg/mL = neutralizing anti-drug-antibodies.

Premium Trace® Assay Principle

For the last decade high sensitive immunoassays required solid phase technologies including extensive washing procedures to reduce background and thereby increase reproducibility and sensitivity. The Premium Trace® assay principle is built upon a new proximity dependent optical assay technology for the detection of specific binding interaction or association between two affinity molecules (antibodies) towards the analyte. One affinity molecule having attached light chemistry and the second affinity molecule having attached an enzyme that can activate the light chemistry on the first affinity molecule. When the two affinity molecules are brought into close proximity through a specific binding event with the analyte a weak flash of light is detected by Premium Trace® light detector (μPMT). The amount of light correlates to the concentration of the particular analyte.

Trace® Premium

trace® Premium is the high-sensitivity version for biomarker detection in the ng/mL- pg/mL range. trace® Premium measures a luminescence signal using a micro light detector (μPMT) in conjunction with Atonomics’ proprietary homogeneous proximity chemistry that takes place in the reaction cuvette.

Trace® Premium - Luminescence detection.
The figure below illustrates the reaction principle.)