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Communication Dans Un Congrès Année : 2019

Towards on-chip EVs separation: a lab-on-chip approach

Résumé

Background and motivation Owe to their complexity in size, origin, membrane markers, there is currently no ideal technology available to relate cell-derived microvesicles (EVs) structure and functions. All currently available methods (flow-cytometry, DLS, TRPS, etc.) have limits in their ability to capture the whole diversity of EVs populations and are not amenable to automation and large-scale analysis of numerous samples. In that context, the overall objective of this study is to develop a miniaturized platform allowing the isolation, fractionation and qualification of microvesicles in µL volume. Methods Based on previous works (1), we propose a lab on chip coupling a hydrodynamic separation module enabling EVs separation according to their size to an affinity-trapping chamber compatible with subsequent SPR and AFM characterization. We designed and fabricated 2.5x2.5cm chips enabling the separation of vesicles at tunable cut-off (150-900nm). The proof-of-concept was done using fluorescent calibration particles (polystyrene and melanin resin nanoparticles) biofunctionnalized with proteins and mimicking EVs in buffer solution. Results Sample was introduced into the chip using a syringe pump or a pressure generator and the filtered sample was simply collected at the chip outlet and redirected towards a biodetection chamber designed as an array of gold plots functionalized with antibodies. We demonstrated the high quality separation of 490nm nanoparticles from 920nm particles in concentrated solution (2.109 to 2.1011 particles/µL). Following sorting step, biosynthetic particles were immunocaptured in a miniaturized module of the NBA platform (2,3) for their subsequent analysis. Conclusion We did the proof-of-concept of on-chip nanoparticles separation and capture demonstrating the ability of miniaturized systems to perform sample fractionation. The tunable properties of the device open the way to a versatile tool for preanalytical steps of EVs, including sorting and concentration, even in complex media.
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Dates et versions

hal-02421493 , version 1 (20-12-2019)

Identifiants

  • HAL Id : hal-02421493 , version 1

Citer

Lyne Pillemont, Daniel Guneysu, Céline Elie-Caille, Wilfrid Boireau, Anne Marie Gué. Towards on-chip EVs separation: a lab-on-chip approach. Annual meeting of the International Society for Extracellular Vesicles (ISEV2019), International Society for Extracellular Vesicles, Apr 2019, Kyoto, Japan. pp.PS04.10. ⟨hal-02421493⟩
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