Analysis of plasma protein adsorption onto PEGylated nanoparticles by complementary methods: 2-DE, CE and Protein Lab-on-chip® system

• Published in: Electrophoresis Vol. 28; no. 13; pp. 2252 - 2261
• Main Authors: Kim, Hyun Ryoung, Andrieux, Karine, Delomenie, Claudine, Chacun, Héléne, Appel, Martine, Desmaële, Didier, Taran, Fréderic, Georgin, Dominique, Couvreur, Patrick, Taverna, Myriam
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01-07-2007; WILEY‐VCH Verlag
• Subjects: 2-D PAGE; Adsorption; Animals; Apolipoprotein E; Apolipoproteins - isolation & purification; Blood Proteins - chemistry; Blood Proteins - isolation & purification; Brain - cytology; Cyanoacrylates - chemistry; Electrophoresis, Capillary - methods; Electrophoresis, Gel, Two-Dimensional - methods; Endothelial Cells - metabolism; Nanoparticles - chemistry; PEGylated nanoparticles; Plasma protein adsorption; Polyethylene Glycols - chemistry; Protein Array Analysis - methods; Protein Lab-on-chip; Rats; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.200600694

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.