title: Analysis of plasma protein adsorption onto PEGylated nanoparticles by complementary methods: 2-DE, CE and Protein Lab-on-chip system.
abstract: The biodistribution of colloidal carriers after their administration in vivo depends on the adsorption of some plasma proteins and apolipoproteins on their surface. Poly(methoxypolyethyleneglycol cyanoacrylate-co-hexadecylcyanoacrylate) (PEG-PHDCA) nanoparticles have demonstrated their capacity to cross the blood-brain barrier (BBB) by a mechanism of endocytosis. In order to clarify this mechanism at the molecular level, proteins and especially apolipoproteins adsorbed at the surface of PEG-PHDCA nanoparticles were analyzed by complementary methods such as CE and Protein Lab-on-chip in comparison with 2-D PAGE as a method of reference. Thus, the ability of those methodologies to identify and quantify human and rat plasma protein adsorption onto PEG-PHDCA nanoparticles and conventional PHDCA nanoparticles was evaluated. The lower adsorption of proteins onto PEG-PHDCA nanoparticles comparatively to PHDCA nanoparticles was evidenced by 2-D PAGE and Protein Lab-on-chip methods. CE allowed the quantification of adsorbed proteins without the requirement of a desorption procedure but failed, in this context, to analyze complex mixtures of proteins. The Protein Lab-on-chip method appeared to be very useful to follow the kinetic of protein adsorption from serum onto nanoparticles; it was complementary to 2-D PAGE which allowed the identification (with a relative quantification) of the adsorbed proteins. The overall results suggest the implication of the apolipoprotein E in the mechanism of passage of PEG-PHDCA nanoparticles through the BBB.
authors.affiliation: Laboratory of Pharmaceutical Technology, University of Paris-Sud, Châtenay-Malabry, France.
authors.forename: Hyun Ryoung
authors.initials: HR
authors.lastname: Kim
authors.forename: Karine
authors.initials: K
authors.lastname: Andrieux
authors.forename: Claudine
authors.initials: C
authors.lastname: Delomenie
authors.forename: Héléne
authors.initials: H
authors.lastname: Chacun
authors.forename: Martine
authors.initials: M
authors.lastname: Appel
authors.forename: Didier
authors.initials: D
authors.lastname: Desmaële
authors.forename: Fréderic
authors.initials: F
authors.lastname: Taran
authors.forename: Dominique
authors.initials: D
authors.lastname: Georgin
authors.forename: Patrick
authors.initials: P
authors.lastname: Couvreur
authors.forename: Myriam
authors.initials: M
authors.lastname: Taverna
mesh_terms: D000327:Adsorption; D000818:Animals; D001053:Apolipoproteins / Q000302:isolation & purification; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification; D001921:Brain / Q000166:cytology; D003487:Cyanoacrylates / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D042783:Endothelial Cells / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001053:Apolipoproteins; D001798:Blood Proteins; D003487:Cyanoacrylates; C504655:poly (methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate); D011092:Polyethylene Glycols