Exosomes isolation tool with nanofluidic concentration device (EXIT)

EXIT's ambition is to develop a state-of-the-art, nanotechnologically driven, analytical platform for the isolation of exosomes from bodily fluids, designed to investigate metabolic, proteomic, immunologic and transcriptional changes in Alzheimer's disease. At present there is no effective therapy to Alzheimer's disease and the mechanism of the disease is not fully understood. Recently, exosomes - nanosize membrane vesicles that are present in all bodily fluids - have been shown to play an important role in the remote communication between cells and to be able to pass the blood-brain barrier. EXIT succeeded to develop a state-of-the-art, nanotechnologically driven, analytical platform for the isolation of exosomes from bodily fluids, designed to investigate metabolic, proteomic, immunologic and transcriptional changes in Alzheimer's disease. The basis of this platform is a novel electro-driven separation and concentration method. EXIT will integrate nanotechnology, transcriptomics, genetics and metabolomics with large existing EU biobanks, enabling the investigation of large sample cohorts of paired cerebrospinal fluid and blood. Within EXIT nanotechnologists, analytical chemists, biologists, geneticists, epidemiologists and clinicians will work together to make the long-awaited breakthrough in exosome isolation and rapidly translate this breakthrough into etiologic, preventive and therapeutic applications. Status: In the remaining few months of the project, we used the remaining resources to complete the processing of the wafers with the final design. The final mask design included the chip designs with highest particles pre-concentration capacity that were selected based on the experimental work from Leiden University. Additionally, we included a new design variant that allows guiding separated particle fractions out of the chip for analysis by other standard analytical equipment, for example mass spectrometry. For fabrication was used the process developed within the project, apart from the final bonding step. Due to the upgrade of bonding equipment the delivery of final chips was delayed by 1.5 months. Additional efforts were put on the development of a new bonding process that yields the same quality of the bond as in previously fabricated chips. The final batch of chips was delivered in April 2021. The coordinator, performing all functional chip testing (unfortunately, testing capacity has been reduced in some periods in 2020/2021 due to the pandemic situation), has desired number of wafers for finalising the experimental work. We have received the confirmation that the chips from the final batch function as they should, and further tests are on-going. When it comes to the dissemination: we are currently working on an article draft describing the development of fabrication process and characterisation of design variants from the first generation of devices. The manuscripts follow experimental work achievements, and when the coordinator completes more tests the possibility of further publications will be discussed. EXIT has participated in the Euronanomed III conference and created a project video. EXIT has been an exciting research project with many possibilities for learning and networking. It became a start of a close collaboration with University of Leiden in Netherlands. Together with partners from EXIT we sent a grant application to a recent Horizon Europe call. In this grant application the technology developed in EXIT plays a central role. We are very proud that the nanofluidic chips fabricated in MiNaLab can be part of many biodiagnostics and life sciences applications requiring exosoms isolation from bodily fluids; and besides other, will be used by a start up from EXIT project and University of Leiden.

Prosjektet EXIT resulterte i utvikling av viktige kompetanser for SINTEF MiNaLab. Blant annet innen nanofluidikk og relaterte fabrikasjonsprossesser, medisinsk diagnostikk, og sensitive elektroseparasjons metoder som er integrerbar på brikke. SINTEF har hatt et fint samarbeid med koordinator og andre partnere som har vist seg i felles publikasjoner, felles søknad i mai 2021 (EIC Pathfinder) og gjensidig interesse for å fortsette samarbeid på dette området. Prossesser og design utviklet gjennom EXIT prosjektet på MiNaLab i samarbeid med andre partnere kan brukes for flere biomedisinske anvendelser. En av resultater fra EXIT prosjektet er en nylig registrert oppstartsbedrift fra Leiden Universitetet som heter EXIT077. På lang sikt kan resultater fra prosjektet ha en potensiell stor betydning både vitenskapelig (nytt teknologisk verktøy for oppdagelse av biomarkører av sykdomer), sosialt (personlig medisink system) og økonomisk (kostbare analytiske metoder).

Around 2 million patients in Europe, Japan and the US are annually diagnosed with Alzheimer?s disease. At present, there is no effective therapy and the mechanism of the disease is not fully understood. Recently, exosomes ? nanosize membrane vesicles - were shown to play a key role in the remote communication between cells and to be able to pass the blood-brain barrier. Exosomes are present in all bodily fluids and, thus, are an appealing resource for diagnostic and etiologic research. EXIT aims to develop a beyond the state-of-the art nanotechnology driven analytical platform for isolation of exosomes from bodily fluids and use this platform to investigate metabolic, proteomic, immunologic and transcriptional changes in Alzheimers disease. The basis of this platform is a novel electrodriven separation and fractionation method - nanochannel induced ion depletion zone isotachophoresis. EXIT will integrate nanotechnology, transcriptomics, genetics and metabolomics with large existing EU biobanks, enabling the investigation of large sample cohorts of paired cerebrospinal fluid and blood. Within EXIT nanotechnologists, analytical chemists, biologists, geneticists, epidemiologists and clinicians will work together to make the long-awaited breakthrough in exosome isolation and rapidly translate this breakthrough into etiologic, preventive and therapeutic applications.