Micro- and Nanofluidic Systems for Nanoparticle Analytics and Cell Membrane Models
Abstract
Lipid nanoparticles (LNPs) serve as carriers for mRNA delivery in a new generation of vaccines, but several challenges remain before this approach can offer broad clinical translation of RNA therapeutics. Within the SSF funded industrial research center FoRmulaEx, we have established an interdisciplinary approach to address some of these challenges. The work in the center is in part focused on the development of micro- and nanofluidic systems combined with advanced surface-sensitive optical microscopy approaches to characterize biological nanoparticles with single nanoparticle resolution, as well as new means to temporally resolve endosomal LNP processing and functional mRNA delivery. The presentation will emphasize our work using these platforms for combined label-free scattering- and fluorescence-based visualization of individual biological nanoparticles,[1] including mRNA containing LNPs. A new means to use these systems to investigate LNP binding to and fusion with endosomal membrane mimics formed on top of nonporous silica[2] will also be presented. Using this approach, we have been able quantify both refractive index and size of individual biological nanoparticles, including precise correlation of these properties with the cargo content measured using specific fluorescence labeling. We have also shown that it is possible to decipher new information regarding the role of protein corona formation on the time-dependent maturation steps that critically unlock cellular LNP uptake and mRNA delivery.[3, 4] These micro- and nanofluidics systems provide detailed information about nanoparticle heterogeneity and how different properties influence their interaction with cellular membranes, thus offering a new means to identify physiochemical characteristics that could potentially facilitates cellular uptake and transfection efficiency. It is thus our hope that this approach will contribute to informed designs of next generation nanocarriers for oligonucleotide delivery.
[1] Sjoberg, M.; Mapar, M.; Armanious, A.; Zhdanov, V. P.; Agnarsson, B.; Hook, F., Time-Resolved and Label-Free Evanescent Light-Scattering Microscopy for Mass Quantification of Protein Binding to Single Lipid Vesicles. Nano Letters 2021, 21 (11), 4622-4628.
[2] Joyce, P.; Joemetsa, S.; Isaksson, S.; Hossain, S.; Larsson, P.; Bergstrom, C.; Hook, F., TIRF Microscopy-Based Monitoring of Drug Permeation Across a Lipid Membrane Supported on Mesoporous Silica. Angew Chem Int Edit2021, 60 (4), 2069-2073.
[3] Gallud, M. J. Munson, K. Liu, A. Idström, H. M. G. Barriga, S. R. Tabaei, N. Aliakbarinodehi, M. Ojansivu, Q. Lubart, J. J. Doutch, M. N. Holme, L. Evenäs, L. Lindfors, M. M. Stevens, A. Collén, A. Sabirsh, F. Höök, E. K. Esbjörner, Time evolution of PEG-shedding and serum protein coronation determines the cell uptake kinetics and delivery of lipid nanoparticle formulated mRNA. bioRxiv 2021: https://doi.org/10.1101/2021.08.20.457104
[4] Aliakbarinodehi, N. Gallud, A. Mapar1, M. Wesén, E. Heydari, S. Jing, Y. Emilsson, G. Liu, K. Sabirsh, A. ZhdanovV.P. Lindfors, L. Esbjörner, E. and Höök F. Interaction Kinetics of Individual mRNA-Containing Lipid Nanoparticles with an Endosomal Membrane Mimic: Dependence on pH, Protein Corona Formation, and Lipoprotein Depletion, in revision
Participate on campus or via Zoom.
On campus: PJ lecture hall
Zoom link: https://gu-se.zoom.us/j/64681043702
Read more about the seminar series
Theoretical Physics Seminar
