Issue 9, 2023

Understanding and optimising the transfection of lipopolyplexes formulated in saline: the effects of peptide and serum

Abstract

Lipopolyplexes (LPDs) are of considerable interest for use as gene delivery vehicles. Here LPDs have been prepared from cationic vesicles (composed of a 1 : 1 molar ratio of DOTMA with the neutral helper lipid, DOPE), singly branched cationic peptides and plasmid DNA. All peptides contained a linker sequence (cleaved by endosomal furin) attached to a targeting sequence selected to bind human airway epithelial cells and mediate gene delivery. The current study investigates the effects of novel Arg-containing cationic peptide sequences on the biophysical and transfection properties of LPDs. Mixed His/Arg cationic peptides were of particular interest, as these sequences have not been previously used in LPD formulations. Lengthening the number of cationic residues in a homopolymer from 6 to 12 in each branch reduced transfection using LPDs, most likely due to increased DNA compaction hindering the release of pDNA within the target cell. Furthermore, LPDs containing mixed Arg-containing peptides, particularly an alternating Arg/His sequence exhibited an increase in transfection, probably because of their optimal ability to complex and subsequently release pDNA. To confer stability in serum, LPDs were prepared in 0.12 M sodium chloride solution (as opposed to the more commonly used water) yielding multilamellar LPDs with very high levels of size reproducibility and DNA protection, especially when compared to the (unilamellar) LPDs formed in water. Significantly for the clinical applications of the LPDs, those prepared in the presence of sodium chloride retained high levels of transfection in the presence of media supplemented with fetal bovine serum. This work therefore represents a significant advance for the optimisation of LPD formulation for gene delivery, under physiologically relevant conditions, in vivo.

Graphical abstract: Understanding and optimising the transfection of lipopolyplexes formulated in saline: the effects of peptide and serum

Supplementary files

Article information

Article type
Paper
Submitted
19 11 2022
Accepted
08 3 2023
First published
24 3 2023
This article is Open Access
Creative Commons BY license

Biomater. Sci., 2023,11, 3335-3353

Understanding and optimising the transfection of lipopolyplexes formulated in saline: the effects of peptide and serum

L. Cui, L. Kudsiova, F. Campbell, D. J. Barlow, H. C. Hailes, A. B. Tabor and M. J. Lawrence, Biomater. Sci., 2023, 11, 3335 DOI: 10.1039/D2BM01905A

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