title: The influence of surface charge on serum protein interaction and cellular uptake: studies with dendritic polyglycerols and dendritic polyglycerol-coated gold nanoparticles.
abstract: Nanoparticles (NPs) have gained huge interest in the medical field, in particular for drug delivery purposes. However, binding of proteins often leads to fast NP uptake and rapid clearance, thereby hampering medical applications. Thus, it is essential to determine and control the bio-nano interface. This study investigated the serum protein interactions of dendritic polyglycerols (dPGs), which are promising drug delivery candidates by means of two dimensional gel electrophoresis (2DE) in combination with mass spectrometry. In order to investigate the influence of surface charge, sulfated (sulfated dendritic polyglycerol [dPGS]) and non-sulfated (dPGOH) surfaces were applied, which were synthesized on a gold core allowing for easier separation from unbound biomolecules through centrifugation. Furthermore, two different sizes for dPGS were included. Although size had only a minor influence, considerable differences were detected in protein affinity for dPGS versus dPGOH surfaces, with dPGOH binding much less proteins. Cellular uptake into human CD14+monocytes was analyzed by flow cytometry, and dPGOH was taken up to a much lower extent compared to dPGS. By using a pull-down approach, possible cellular interaction partners of serum pre-incubated dPGS-Au20 NPs from the membrane fraction of THP-1 cells could be identified such as for instance the transferrin receptor or an integrin. Clathrin-mediated endocytosis was further investigated using chlorpromazine as an inhibitor, which resulted in a 50% decrease of the cellular uptake of dPGS. This study could confirm the influence of surface charge on protein interactions and cellular uptake of dPGS. Furthermore, the approach allowed for the identification of possible uptake receptors and insights into the uptake mechanism.
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Berlin, Germany.
authors.forename: Tony
authors.initials: T
authors.lastname: Bewersdorff
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Vonnemann
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Berlin, Germany.
authors.forename: Asiye
authors.initials: A
authors.lastname: Kanik
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Haag
authors.affiliation: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), Berlin, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Haase
mesh_terms: D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000494:pharmacology; D016503:Drug Delivery Systems / Q000379:methods; D004705:Endocytosis / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods; D005990:Glycerol / Q000737:chemistry / Q000493:pharmacokinetics*; D006046:Gold / Q000737:chemistry; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D011108:Polymers / Q000737:chemistry / Q000493:pharmacokinetics*; D000066970:Protein Corona / Q000737:chemistry; D013431:Sulfates / Q000737:chemistry
chemical_list: D001798:Blood Proteins; D018950:Lipopolysaccharide Receptors; D011108:Polymers; D000066970:Protein Corona; D013431:Sulfates; C043941:polyglycerol; D006046:Gold; D005990:Glycerol; D002746:Chlorpromazine
keywords: cellular uptake; clathrin-mediated endocytosis; protein corona; sulfated dendritic polyglycerols; uptake receptors