Bohórquez, Ana C.

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Testing of centrifugal filter devices as a tool to quantify protein-nanoparticle interactions using UV/Vis measurements
(2012) Bohórquez, Ana C.; Latorre-Esteves, Magda; College of Engineering; Rinaldi, Carlos; Torres-Lugo, Madeline; Department of Chemical Engineering; Monroig Saltar, Francisco M.
Centrifugal Filter Devices (CFD) has been tested for the quantification of protein adsorption onto the surface of magnetic nanoparticles coated with biocompatible polymers. Using a phosphate buffered saline wash protocol, the bicinchoninic acid assay (BCA), and mass balance calculations, the amount of protein adsorbed was calculated. The iron in magnetic cores interferes with the BCA assay because it remains in the supernatant after centrifugation. CFD was considered an attractive alternative, because in principle it could retain colloidally stable nanoparticles, leaving unbound protein to be quantified in the filtrate. Magnetic nanoparticles were synthesized by co-precipitation and grafted with Carboxylmethly Dextran or silanized polyethylene glycol. Proteins used for this study include anionic and cationic proteins: Bovine Serum Albumin and Lysozyme from Hen Egg White, respectively. In conclusion, CFDs were not suitable for the study of protein-nanoparticle interactions because membrane fouling and concentration polarization were encountered; critical factors that compromised measurement accuracy.