Design and validation of sampling devices for ocular surface and fluids

Personalized medicine is a growing field in which physicians and biologists use diagnostic tests to identify specific biological markers, which will determine medical treatments and procedures for each patient. Merging the researcher information with medical records, allows clinicians to develop targeted treatment and prevention plans (Offit K, 2011; Wilson B, 2015). Various scientific publications describe extensively the personalized medicine, health information management, biomarker discovery and targeted therapies (Zhang L, 2015). The market of molecular diagnostics devices is growing fast, nearly 4000 new diagnostic tests have been introduced only in 2015 (Pritchard DE, 2017). Equal for the molecular therapeutics market, nearly 28% of all the medicines the United States Food and Drugs Administration (FDA) approved in 2015 were personalized medicines (https://www.fda.gov/Drugs/DevelopApprovalProcess), and a recent study sponsored by the Personalized Medicine Coalition (PMC) demonstrated that 42% of all medicines and 73% of cancer medicines in development are potential personalized medicines (Pritchard DE, 2017). To realize a personalized approach new technology that endorses a person's biology, like DNA, RNA, or protein, to confirm a possible disease is required (Vogenberg FR, 2010). In human medicine, and in particular in Ophthalmology, suitable cellular markers can be used to recognize corneal and conjunctival pathologies; their use is very useful for the diagnosis of ocular diseases. These markers provide great help both from a diagnostic point of view and from a therapeutic point of view. This type of investigation would in fact determine benefits from the etiopathogenetic point of view, clinical, diagnostic and prognostic for the pathology in consideration. Hence, the main aim of my Ph.D. was to simplify the discovery of specific eye disorder biomarkers, through the development of new sampling and testing devices, that allow the identification of a specific biomarker, correlated to a specific pathology, in office. Therefore, my functional intent for the present thesis as shown in scheme 1 was to undergo the simplification of biomarker discovery, through the design and development of three different devices: • First a new sampling device, that incorporates a nitrocellulose membrane, that permits a fast and reliable imprinting and analysis of the ocular surface, as compared to other old sampling techniques, referring to biochemical and molecular analyses in order to identify and have an overview of the enormous field of the Biomarkers. • Second a solubilizing device, that attends to solubilize the proteins and nucleic acids contained in biological fluids, or from the nitrocellulose membrane of the new sampling device, obtaining therefore samples immediately available for analysis. • Third a biosensor, to facilitate the research of biomarkers of interest, I have participated in the development of a biosensor, for an already patented device, that will complete it, giving a quantifiable biomarker response, in collaboration with the ISM, IMM IFT institutes of the CNR of Tor Vergata. This last device will give an immediate and quantitative response of a biomarker, solubilized in the second device in which both the biological fluids and the nitrocellulose membrane of the first device can be inserted, contributing to pursuing, therefore, the same goal, or the easiest identification of one or more specific pathology markers.