R&D of innovative sensors for health

1. Biomedical, in vivo and in vitro diagnostic R&D of innovative sensors for health DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it Elettra-Sincrotrone Trieste UNIVR ThunderNILS.r.l. ICGEB Trieste UNITS 26 September 2018

2. 26 September 2018 The project is divided into 4 different research areas, each of them is identified by an acronym for convenience of reference below. The areas are: Study and testing of capacitively-based biosensor systems for detection of biomarkers in serum. Acronym BSR. Chronic myeloid leukemia - Nano-Electrode Array. Acronym CML - NEA Human Papilloma Virus - Nano-Electrode Array. Acronym HPV - NEA. Biosensor for Unbound Bilirubin. Acronym BBL. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

3. 26 September 2018 BBL – Biosensor for Unbound Bilirubin • State of art • Bilirubin (BR) fractions in plasma: • - Conjugated BR (direct) • Unconjugated BR (indirect) • The Unconjugated fraction accounts for: • Unbound bilirubin • Bilirubin bound to albumin • Current methods for BR detection mainly focus on the Conjugate (direct) fraction and Total BR, hiding the real levels of Unbound BR. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

4. 26 September 2018 BBL – Biosensor for Unbound Bilirubin Diagnostic value Unbound bilirubin levels in plasma are recently reported as companion diagnostic for chronic pathologies, over the role as marker of hepatic function. Project aim Starting from a developed biopolimeric fluorescent biosensor for unbound bilirubin (BBL) immobilized on a 2D scaffold in solid phase, the project aims to: • Scale-up of BBL protein production, from E. Coli culture; • 2D scaffold characterization for BBL immobilization; • Performance evaluation of immobilized BBL for unbound BR detection, both in standard solutions and in real biological samples. • The BR dependent fluorescence of this solid phase biosensor can be easily determined in 96well plates with microplates reader. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

5. 26 September 2018 BSR Project aim CIRculating biomarkers Electrochemical Impedance sensors(CIRCEI) will be used for detection of phosphorilation state of a peptide biosensor for CML. P P P P P P P P P Enzyme phosphorilation Signal acquiring correlated to enzyme phosphorilation activity Peptide biosensor linker surface DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

6. 26 September 2018 CML-NEA (NEA biosensors for Bcr-Abl activity monitoring) Pasic I, Lipton JH. Leuk Res. 2017 Apr. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

7. 26 September 2018 CML-NEA (NEA biosensors for Bcr-Abl activity monitoring) • Develop a platform useful for monitoring the activity of the chimeric kinase ABL and the inhibition of tyrosine kinase inhibitor drugs. • Early identification of effective treatments for CML. • Transform the device to point of care (PoC) test based of NEA biosensors. Kim YJ., et al. Biosens Bioelectron. 2018 Oct 15.  DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

8. 26 September 2018 E6/E7 PHOSPHORILATES: ROLE IN TUMOR PROGRESSION DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

9. 26 September 2018 DETECTION OF PROGRAMMING MARKERS OF HPV INFECTION (PROTEIN E6-E7) THROUGH ELECTROCHEMICAL SENSORS Assembled prototype of electrochemical nanosensor based on the Nanoelectrode arrays (NEA); detection scheme with immobilization of the target protein, primary antibody and secondary antibody with enzymatic label and diagram of the electrochemical measurement cell for the measurement. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

10. 26 September 2018 Conclusions These innovative sensors for health can have an impact on the market. BBL Bilirubinemia is one of the routine blood chemistry parameters. The biosensor will allow an extremely more precise measurement and the determination of the free and unbounded fraction, which is currently not analytically accessible. By virtue of its use as a companion diagnostic for personalized medicine (or precision medicine), this product can be used primarily for screening healthy population. It can be assumed that ten million tests/year are used. BSR Miniaturized capacitive devices have a potential very broad market and can be used for cancer and tumor recurrence screening, cardiovascular disease screening and monitoring of the efficacy of anti-tumor therapy. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

11. 26 September 2018 Conclusions CML – NEA The device will be used to predict the response to drugs and determine therapy. From this use the potential market can be estimated from the incidence of CML equal to about 10-15 x 10^6 cases/year in the general population with a market of 140000 tests/year in the European Union only . HPV – NEA This diagnostic test could be developed by exploiting the results from industrial research in this field of application has features allowing to aim, after adequate validation at the HPV screening market, to be used as an alternative test for the current Pap test in the early stage of cervical cancer prevention. DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it

12. 26 September 2018 DOTT. DINO PALADIN Area Scienze Park Padriciano 99 34149 - Trieste Incubatori FVG BIC Via Flavia 23 34148 - Trieste www.dinopaladin.it dinopaladin@dinopaladin.it