PuSH - Publikationsserver des Helmholtz Zentrums München: Protein corona investigations of polyplexes with varying hydrophobicity - From method development to in vitro studies.

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Hartl, N.* ; Jürgens, D.C.* ; Carneiro, S.* ; König, A.-C. ; Xiao, X.* ; Liu, R.* ; Hauck, S.M. ; Merkel, O.M.*

Protein corona investigations of polyplexes with varying hydrophobicity - From method development to in vitro studies.

Int. J. Pharm. 643:123257 (2023)

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In the field of non-viral drug delivery, polyplexes (PXs) represent an advanced investigated and highly promising tool for the delivery of nucleic acids. Upon encountering physiological fluids, they adsorb biological molecules to form a protein corona (PC), that influence PXs biodistribution, transfection efficiencies and targeting abilities. In an effort to understand protein - PX interactions and the effect of PX material on corona composition, we utilized cationic branched 10 kDa polyethyleneimine (b-PEI) and a hydrophobically modified nylon-3 polymer (NM_0.2/CP_0.8) within this study to develop appropriate methods for PC investigations. A centrifugation procedure for isolating hard corona - PX complexes (PCPXs) from soft corona proteins after incubating the PXs in fetal bovine serum (FBS) for PC formation was successfully optimized and the identification of proteins by a liquid chromatography-tandem mass spectrometry (LC-MS-MS) method clearly demonstrated that the PC composition is affected by the underlying PXs material. With regard to especially interesting functional proteins, which might be able to induce active targeting effects, several candidates could be detected on b-PEI and NM_0.2/CP_0.8 PXs. These results are of high interest to better understand how the design of PXs impacts the PC composition and subsequently PCPXs-cell interactions to enable precise adjustment of PXs for targeted drug delivery.

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