Analysis of plasma protein adsorption onto PEGylated nanoparticles by complementary methods: 2-DE, CE and Protein Lab-on-chip system.
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.
Kim
Hyun Ryoung
HR
Laboratory of Pharmaceutical Technology, University of Paris-Sud, Châtenay-Malabry, France.
Andrieux
Karine
K
Delomenie
Claudine
C
Chacun
Héléne
H
Appel
Martine
M
Desmaële
Didier
D
Taran
Fréderic
F
Georgin
Dominique
D
Couvreur
Patrick
P
Taverna
Myriam
M
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
D001053:Apolipoproteins; D001798:Blood Proteins; D003487:Cyanoacrylates; C504655:poly (methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate); D011092:Polyethylene Glycols