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. 2023 Jan:47:102626.
doi: 10.1016/j.nano.2022.102626. Epub 2022 Nov 7.

Direct quantification of cytosolic delivery of drug nanocarriers using FlAsH-EDT2

R Rotem  1 , J A Bertolini  2 , L Salvioni  2 , L Barbieri  2 , M A Rizzuto  2 , V Tinelli  2 , A Gori  3 , S Adams  4 , M Colombo  2 , D Prosperi  5
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Direct quantification of cytosolic delivery of drug nanocarriers using FlAsH-EDT2

R Rotem et al. Nanomedicine. 2023 Jan.

Abstract

The delivery of therapeutics across the cell membrane and into the cytoplasm is a major challenge that limits the development of new therapies. This challenge is compounded by the lack of a general assay for cytosolic delivery. Here we develop this assay based on the pro-fluorophore CrAsH-EDT2, and provide cytosolic penetration results for a variety of drug delivery agents (polyethyleneimine, poly-arginine, Ferritin, poly [maleic anhydride-alt-isobutene] grafted with dodecylamine, and cationic liposomes) into HeLa and T98G cells. Our results show that this method can be widely applicable to different cells and drug delivery agents, and yield statistically robust results. We later use this method to optimize and improve a model drug delivery agent's (Ferritin) cytosolic penetration.

Keywords: Cell penetration; Drug delivery; FlAsH-EDT2; Flow cytometry; Quantification method.

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Conflict of interest statement

Declaration of competing interest The authors declare that they have no competing interests.

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