| Autor: |
Biswas A; Institute of Technology, University of Tartu, 1 Nooruse Street, 50411 Tartu, Estonia., Maloverjan M; Institute of Technology, University of Tartu, 1 Nooruse Street, 50411 Tartu, Estonia., Padari K; Institute of Molecular and Cell Biology, University of Tartu, 23b Riia Street, 51010 Tartu, Estonia., Abroi A; Institute of Technology, University of Tartu, 1 Nooruse Street, 50411 Tartu, Estonia., Rätsep M; Institute of Physics, University of Tartu, 1 Wilhelm Ostwaldi, 51014 Tartu, Estonia., Wärmländer SKTS; Chemistry Section, Stockholm University, Arrhenius Laboratories, SE-10691 Stockholm, Sweden.; CellPept Sweden AB, Kvarngatan 10B, SE-11847 Stockholm, Sweden., Jarvet J; Chemistry Section, Stockholm University, Arrhenius Laboratories, SE-10691 Stockholm, Sweden.; CellPept Sweden AB, Kvarngatan 10B, SE-11847 Stockholm, Sweden., Gräslund A; Chemistry Section, Stockholm University, Arrhenius Laboratories, SE-10691 Stockholm, Sweden.; CellPept Sweden AB, Kvarngatan 10B, SE-11847 Stockholm, Sweden., Kisand V; Institute of Physics, University of Tartu, 1 Wilhelm Ostwaldi, 51014 Tartu, Estonia., Lõhmus R; Institute of Physics, University of Tartu, 1 Wilhelm Ostwaldi, 51014 Tartu, Estonia., Pooga M; Institute of Technology, University of Tartu, 1 Nooruse Street, 50411 Tartu, Estonia. |
| Abstrakt: |
Cell-penetrating peptides (CPPs) are highly promising transfection agents that can deliver various compounds into living cells, including nucleic acids (NAs). Positively charged CPPs can form non-covalent complexes with negatively charged NAs, enabling simple and time-efficient nanoparticle preparation. However, as CPPs have substantially different chemical and physical properties, their complexation with the cargo and characteristics of the resulting nanoparticles largely depends on the properties of the surrounding environment, i.e., solution. Here, we show that the solvent used for the initial dissolving of a CPP determines the properties of the resulting CPP particles formed in an aqueous solution, including the activity and toxicity of the CPP-NA complexes. Using different biophysical methods such as dynamic light scattering (DLS), atomic force microscopy (AFM), transmission and scanning electron microscopy (TEM and SEM), we show that PepFect14 (PF14), a cationic amphipathic CPP, forms spherical particles of uniform size when dissolved in organic solvents, such as ethanol and DMSO. Water-dissolved PF14, however, tends to form micelles and non-uniform aggregates. When dissolved in organic solvents, PF14 retains its α-helical conformation and biological activity in cell culture conditions without any increase in cytotoxicity. Altogether, our results indicate that by using a solvent that matches the chemical nature of the CPP, the properties of the peptide-cargo particles can be tuned in the desired way. This can be of critical importance for in vivo applications, where CPP particles that are too large, non-uniform, or prone to aggregation may induce severe consequences. |
