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. Author manuscript; available in PMC: 2022 May 9.
Published in final edited form as: Nat Nanotechnol. 2022 Feb 7;17(3):310–318. doi: 10.1038/s41565-021-01030-y

Figure 1.

Figure 1.

Characterizing species-agnostic nanoparticle delivery screening (SANDS). (a) LNP-N is formulated to carry mRNA encoding aVHH as well as DNA barcode N. (b) Stable LNPs are pooled and injected into mice; aVHH+ cells are later isolated by FACS, and the barcodes within the cells are identified using next-generation sequencing. (c) Anchored VHH (aVHH) was selected as a reporter since it is embedded into the cell bilayer and can be quantified using antibodies against VHH. (d) aVHH protein expression, measured by immunofluorescent staining, 24 hours after A549 cells were transfected with aVHH mRNA. (e) aVHH protein expression, measured by flow cytometry, in Fa2N-4 human and AML-12 mouse hepatocytes 24 hours after transfection with aVHH mRNA. (f) The percentage of aVHH+ hepatocytes (i.e., CD31−CD45−aVHH+) at different timepoints following treatment with LNPs carrying 1 mg/kg aVHH mRNA. N=3/group, average +/− SEM. (g) The combinatorial design of 144 chemically distinct LNPs formulated carrying aVHH and DNA barcodes. (h) Hydrodynamic diameters of 89 LNPs that were deemed stable enough to pool together, as well as the diameter of the pooled LNP control. (i-l) Hydrodynamic diameter of the 89 LNPs plotted as a function of LNP chemical property. N=3/group, average +/− STD.