title: Quantitative proteomics analysis of adsorbed plasma proteins classifies nanoparticles with different surface properties and size.
abstract: Nanoparticle biological activity, biocompatibility and fate can be directly affected by layers of readily adsorbed host proteins in biofluids. Here, we report a study on the interactions between human blood plasma proteins and nanoparticles with a controlled systematic variation of properties using (18)O-labeling and LC-MS-based quantitative proteomics. We developed a novel protocol to both simplify isolation of nanoparticle bound proteins and improve reproducibility. LC-MS analysis identified and quantified 88 human plasma proteins associated with polystyrene nanoparticles consisting of three different surface chemistries and two sizes, as well as, for four different exposure times (for a total of 24 different samples). Quantitative comparison of relative protein abundances was achieved by spiking an (18)O-labeled "universal" reference into each individually processed unlabeled sample as an internal standard, enabling simultaneous application of both label-free and isotopic labeling quantification across the entire sample set. Clustering analysis of the quantitative proteomics data resulted in distinctive patterns that classified the nanoparticles based on their surface properties and size. In addition, temporal data indicated that the formation of the stable protein corona was at equilibrium within 5 min. The comprehensive quantitative proteomics results obtained in this study provide rich data for computational modeling and have potential implications towards predicting nanoparticle biocompatibility.
authors.affiliation: Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors.forename: Haizhen
authors.initials: H
authors.lastname: Zhang
authors.forename: Kristin E
authors.initials: KE
authors.lastname: Burnum
authors.forename: Maria L
authors.initials: ML
authors.lastname: Luna
authors.forename: Brianne O
authors.initials: BO
authors.lastname: Petritis
authors.forename: Jong-Seo
authors.initials: JS
authors.lastname: Kim
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Qian
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Moore
authors.forename: Alejandro
authors.initials: A
authors.lastname: Heredia-Langner
authors.forename: Bobbie-Jo M
authors.initials: BJ
authors.lastname: Webb-Robertson
authors.forename: Brian D
authors.initials: BD
authors.lastname: Thrall
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.forename: Joel G
authors.initials: JG
authors.lastname: Pounds
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
mesh_terms: D000327:Adsorption; D000704:Analysis of Variance; D001798:Blood Proteins / Q000032:analysis* / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D016000:Cluster Analysis; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D011137:Polystyrenes / Q000737:chemistry; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D013499:Surface Properties
chemical_list: D001798:Blood Proteins; D011137:Polystyrenes