The ELIDIS project was structured into four work packages (WPs), spanning technical validation (WP1), prototype design and manufacturing (WP2), IP portfolio development (WP3), and market study and business planning (WP4). Below we describe the main activities carried out and the key results achieved over the 18-month project duration.
WP1 — Technical Validation
In the first phase of the project, we focused on the full analytical characterization of the ELIDIS assay for the two model targets. We synthesized and optimized antigen-conjugated DNA strands and enzyme (glucose oxidase)-DNA hybrid constructs using click chemistry and HPLC/gel electrophoresis purification protocols. Both direct (for cetuximab) and competitive (for EGFR) assay formats were systematically optimized on disposable 8- and 24-electrode screen-printed arrays produced in-house using our DEK 248 screen printer.
For Cetuximab detection, the ELIDIS platform achieved a limit of detection (LOD) of 1×10^(-12)M, with a relative standard deviation (RSD) below 12% in 80% serum. In the presence of non-specific antibodies, the recorded signals were indistinguishable from the background, demonstrating the high specificity of the assay.
A critical milestone of WP1 was the head-to-head comparison of the ELIDIS assay with commercially available ELISA kits. For trastuzumab, we compared against the Abcam Herceptin ELISA Kit (sensitivity: 11 ng/mL, price: ~1,650 €/96 tests) and the Creative Diagnostics Human Herceptin ELISA Kit (0.24 ng/mL, ~1,400 €). For EGFR, comparisons were made with the ThermoFisher EGFR Full-length Human ELISA Kit (0.2 ng/mL, ~800 €) and the Abcam Human EGFR ELISA Kit (4 pg/mL, ~760 €).
The performance of ELIDIS was compared with that of a commercial ELISA kit (Erbitux from Assay Genie) for the detection of Cetuximab. ELIDIS demonstrated superior analytical sensitivity, achieving an LOD of 1×10^(-12)M compared with 6×10^(-10)M for the commercial ELISA kit. In addition, ELIDIS required fewer experimental steps, including only a single washing step, while maintaining a comparable overall assay time. Recovery studies were conducted using blank serum samples spiked with Cetuximab at concentrations ranging from 10^(-12)to 10^(-7)M and analysed using both the ELIDIS platform and the commercial ELISA assay. Comparable recovery values were obtained for ELIDIS and ELISA within the concentration range of 10^(-8)to 10^(-7)M. Notably, the commercial ELISA kit did not provide quantitative measurements below 10^(-9)M, whereas ELIDIS maintained sensitivity down to 10^(-12)M.
We also initiated development of the instrument-free detection mode by starting the development of a dedicated app/software to enable chronoamperometric control of the electrode array via USB-C connection to a smartphone or laptop, without the need of the PalmSense portable potentiostat.
WP2 — Prototype Design and Manufacturing Plan
Following technical validation, we developed a prototype ELIDIS kit in a disposable format. All assay components (DNA strands, enzyme conjugates) were lyophilized (freeze-dried) directly into disposable Eppendorf tubes, enabling a simple add-sample-and-measure workflow. A 24-electrode screen-printed array was integrated into the kit, and the full assay workflow — from sample addition to electrochemical readout — was demonstrated.
A manufacturing plan was also prepared, describing the reagent preparation, quality control steps, and electrode fabrication workflow in a format suitable for transfer to a contract manufacturer or industrial partner.
WP3 — IP Portfolio
The Italian patent application no. 102023000014028, covering the general principle of the ELIDIS technology, was filed prior to the project start and received a very positive patent search report in February 2024. During the ELIDIS PoC project, the international (PCT) extension of this patent was pursued ahead of the April 2025 deadline.
An IP strategy document was also developed in collaboration with the Innovation and Incubation Center of INBB, identifying the strongest patentable aspects of the kit format, the lyophilized component design, and the multiplexed detection capability.
WP4 — Market Study and Business Plan
A detailed market analysis was conducted covering the immunoassay market, competitive landscape, distribution channels, and end-user segments. Contacts were established with key potential partners including Merck, Dynamic Biosensors, Ulisse Biomed, and Biosearch Technology.
