All of the four ESRs have been successfully recruited and have been enrolled in Medical Biology PhD Program of Paris-Lodron University Salzburg, one of academic partners, and in Molecular Life Sciences PhD Program of “Luigi Vanvitelli” University Caserta, Italy. Due to a huge amendment that had to be implemented at the very beginning of the project and restrictions due to Covid-19 pandemic, the ESRs have started their research activities with some delays and difficulties. However, they have collected promising results on their individual projects. The research activities mainly addressed: 1.Interaction of endotoxin with biological molecules; 2. Interaction of endotoxin with particulate matter; 3. NP functionalisation for optimal endotoxin capture/scavenging in different media; 4. Synthesis of detection probes based on Fluorescence or Raman signal for optimal detection and quantitative signalling of endotoxin presence. They addressed the interaction of endotoxin from different bacteria with cells, such as human primary innate immune cells, human monocytic cell lines transfected with NFkB luciferase to receive a luminescence signal depending on LPS induced signaling. New method based on magnetic microbeads has been set up to uncover novel LPS binding molecules from promising sources such as cell lysate. Biological molecules, especially new antibodies obtained through Phage and Yeast Display, have been identified as potential new biosensors and for NP optimization/conjugation. Conjugation conditions (molecule/nanoparticle ratio, buffer, time and temperature) have been optimised in order to establish the stability of gold and polystyrene nanoparticles complexed with newly identified through Phage Display (scFv-Fc #2) or selected (Anti Lipid A polyclonal antibody) antibodies. Total Internal Reflection Fluorescence (TIRF) or Surface Enhanced Raman Scattering (SERS) configuration were established and silver nanoparticles deposited on glass surfaces has been identified to be used in the development of LPS immunoassay. Moreover, Immuno Magnetic Assay for the LPS detection developed at Blusense Diagnostics has been implemented by using iron oxide magnetic nanoparticles in order to set up an Innovative optomagnetic assay based on the recognition of LPS by new identified aforementioned antibodies and on the agglutination of magnetic nanoparticles.
The ENDONANO ESRs performed numerous dissemination activities (such as preparing publications and taking part in international scientific meetings or in public events such as the European Biotech Week and “Italia: l'arte della scienza”, and INDUSTRY DAYS and AUTOMOTIVE HUNGARY) and attended courses on topics such as “Responsible Research and Innovation”, “Communication to the general public and risk communication”, and “IP, regulatory aspects and entrepreneurship”. The latter activities helped building their scientific background and increasing their opportunities for a future, successful career in science. Therefore, the project has achieved significant scientific advances and has successfully trained four young scientist in the field of nanomaterials or nano-medical devices through a truly interdisciplinary approach.
