The project has started with the identification of end users requirements and technical specifications of ULTRAPLACAD final industrial prototype. Efficient synthetic probes for the detection of nucleic acid sequences have been designed and synthesized and procedures for the needed chemical modification of nanostructures have been selected and tested. Nanoparticle-enhanced assays that operate with attomolar sensitivity in detecting DNA and femtomolar sensitivity in detecting microRNAs have been developed. PEFSI detection of a-TAAs is performed by attaching specifically designed and synthesized thermoresponsive polymers on PEFSI nanostructures.
The industrial prototype of a bimodal reader for NESPRI and PEFSI detection has been developed and tested thanks to the knowledge acquired by designing and fabricating NESPRI and PEFSI laboratory prototypes. A leaflet describing the industrial prototype was prepared to attract potential customers.
Animal models of liquid biopsy have been developed and retrospective cohorts of colorectal carcinoma patient plasma samples have been recruited. An internal mini-bank comprising tumor tissue, plasma, and purified analytes was organized and samples shared among partners.
The process for the high-volume fabrication of nanostructured disposable chips has been developed and demonstrated. The microfluidics part of the disposable chip has been produced by demonstrating the high-volume manufacturing capability. The integration of the nanostructured optical surface and microfluidics part in large-volume production has also been demonstrated. Protocols for the PCR-free detection of circulating DNAs and microRNAs from human plasma have been defined.
The ULTRAPLACAd industrial prototype has been installed at the Laboratory of Clinical Pathology of the Italian National Cancer Institute (Rome) and a public demonstration was given during the ULTRAPLACAD end-user workshop.
The project results were made available to the scientific community through 19 Open Access peer review publications and to broad public through the project website and YouTube video (
https://www.youtube.com/watch?v=88n3IRsWTm8(si apre in una nuova finestra)). Exploitation strategies were discussed and defined. They include participation to new EU funded projects (short term), commercialize the platform (long term) and agreement with companies outside the consortium (long term).
To summarize, the main results achieved are:
- NESPRI and PEFSI nanostructure geometries have been identified;
- Probes for the selected all-RAS mutations and microRNAs have been synthesized;
- Candidate antigenic peptides for a-TAA detection have been defined;
- Attomolar SPRI detection of all-RAS mutations and femtomolar detection of microRNAs have been demonstrated;
- The high-volume fabrication of nanostructured disposable chips has been demonstrated;
- The ULTRAPLACAD industrial prototype, based on the NESPRI and PEFSI readers lab prototypes, has been fabricated, tested and public demonstration given.
