ViroZoom

The aim of this project is to develop a nanopore-based technology for analysing viruses at the level of the individual particle. Our aim is to build a multimodal characterisation platform, calibrated to measure several viral properties that give particles their infectious potential. Our ambition in this project is to test our approach in relevant experimental contexts but also to address well-defined challenges that are not covered by existing technologies, in particular for diagnostic purposes and also for the bioproduction of viral vectors. Our method is based on nanopore technology coupled with optical detection to enable real-time monitoring of virus transport. This configuration enables precise quantification of viral concentration. In addition, the identification of virus traffic jams in the nanopores, coupled with modelling, provides access to dynamic virus interaction parameters. We have shown that these parameters constitute a unique signature for each viral type and even sub-type. On the basis of promising preliminary results, this project aims to establish a characterisation method on real samples that will enable us to measure the surface state or the presence of the genome. The first aspect of the project is to design a miniaturised, automated prototype suitable for routine use in microbiological high-security laboratories (P2/P3). To this end, it will be adapted to enable the acquisition of two colours simultaneously. Image analysis will be automated by adapting existing algorithms initially developed for super-resolution microscopy or for segmentation using Deep Learning. The second part of the project is devoted to the development of diagnostic tools. We will be focusing on respiratory viruses (InflV, HSV, SC2), which represent a major challenge for global health due to their extent and variability. Following preliminary data on negative matrices, we will apply our methodology to patient samples by developing a sample pre-treatment system. We will then differentiate between infectious and non-infectious particles and characterise viral surface glycoproteins. The third axis focuses on the qualification of viral bioproductions. We will use an effective model for industrial-scale vaccine production: the baculovirus/insect cell system to produce Sars-Cov-2 virus-like particles. The main objectives are to quantify undesirable co-secreted particles and to analyse the glycosylation of viral surface proteins. Our approach will make it possible to optimise bioproduction parameters by carrying out quality control of production batches. Our consortium brings together complementary expertise in single-molecule physics, virology, human viral infectiology and vaccine production. The ambition of this project is to improve viral analysis by providing a high-throughput methodology applicable to a wide range of viruses encountered in biotechnology or as human pathogens.