title: Comparative examination of adsorption of serum proteins on HSA- and PLGA-based nanoparticles using SDS-PAGE and LC-MS.
abstract: The behavior of nanosized drug carrier systems under cell culture conditions and therefore also the destiny in the body are highly influenced by the protein corona, which is formed upon entering a biological environment. Some of the adsorbed proteins, named opsonins, lead to a shortened plasma circulation half-life of the nanoparticles. Others are attributed to promote the transport of nanoparticles into other compartments of the body, just to mention two examples. Hence, detailed knowledge concerning the composition of the protein corona is of great importance. The aim of this work was to investigate the influence of the nanoparticle starting material and the surface modification on the composition of the adsorbed serum proteins in a cell culture environment. Therefore, positively charged nanoparticles based on the biodegradable polymer poly(dl-lactide-co-glycolide) (PLGA) stabilized with didodecyldimethylammonium bromide (DMAB) and negatively charged nanoparticles based on human serum albumin (HSA) were prepared and modified with hydrophilic polymers. By incubating the nanoparticles with fetal bovine serum (FBS) the adsorption of serum proteins on the colloidal system was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) a semi-quantitative analysis of the protein corona was performed and after enzymatic in-solution-digestion the adsorbed proteins were identified using high resolution LC-MS. Our study accentuates the influence of the core material, surface charge, and surface modification on the amount and nature of the adsorbed proteins. The combination of SDS-PAGE and LC-MS turns out to be a simple and reliable method to investigate the protein corona of nanoparticles.
authors: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstraße 48, 48149 Münster, Germany.
authors: R
authors: R
authors: Gossmann
authors: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstraße 48, 48149 Münster, Germany.
authors: E
authors: E
authors: Fahrländer
authors: Institute of Food Chemistry, University of Muenster, Corrensstraße 45, 48149 Münster, Germany.
authors: M
authors: M
authors: Hummel
authors: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstraße 48, 48149 Münster, Germany.
authors: D
authors: D
authors: Mulac
authors: Institute of Food Chemistry, University of Muenster, Corrensstraße 45, 48149 Münster, Germany.
authors: J
authors: J
authors: Brockmeyer
authors: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstraße 48, 48149 Münster, Germany. Electronic address: k.langer@wwu.de.
authors: K
authors: K
authors: Langer
mesh_terms: D000327:Adsorption; D002851:Chromatography, High Pressure Liquid; D003102:Colloids; D004337:Drug Carriers; D004591:Electrophoresis, Polyacrylamide Gel; D057927:Hydrophobic and Hydrophilic Interactions; D019344:Lactic Acid / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry; D050997:Nanomedicine / Q000379:methods*; D053758:Nanoparticles; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000378:metabolism; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011485:Protein Binding; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D000075462:Serum Albumin, Human; D052616:Solid Phase Extraction; D013499:Surface Properties; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: C422455:ALB protein, human; D003102:Colloids; D004337:Drug Carriers; D000644:Quaternary Ammonium Compounds; D012709:Serum Albumin; C046112:didodecyldimethylammonium; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D000075462:Serum Albumin, Human
keywords: Didodecyldimethylammonium bromide; Human serum albumin; LC–MS; Nanoparticles; Poly(ethylene glycol); Poly(lactic-co-glycolic acid); Polyvinyl alcohol; Protein adsorption; SDS–PAGE