Probing nanoparticle--protein interaction by capillary electrophoresis.
Understanding nanoparticle-protein interaction could help in promoting applications of nanoparticles in the biomedical fields and reducing/preventing possible adverse effects to the biological systems caused by nanoparticles. Quantitative measurement of the biophysical parameters of nanoparticle-protein interaction will improve such understanding, which could be conveniently performed by capillary electrophoresis (CE) as demonstrated in the present study. Two interaction situations were identified. Stable nanoparticle-protein complexes were resolved from the free nanoparticles and the proteins by capillary zone electrophoresis (CZE). Transient complexes with fast association/dissociation rates showed distinct mobility change from the free nanoparticles in affinity capillary electrophoresis (ACE). Interactions of bovine serum albumin (BSA) with the Fe(3)O(4) nanoparticles (average diameters of 8 and 10 nm) and with the Au nanoparticles (average diameters of 5 and 10 nm) displayed slow and fast binding kinetics, respectively. Using the Hill equation, we could calculate the dissociation constants (K(D)) and cooperativity coefficients (n). Impacts on nanoparticle-protein interaction from the physicochemical properties of nanoparticles and the incubation buffer were evaluated on the basis of the K(D) and n values to interpret the interaction driving forces. Our study demonstrated the high simplicity and flexibility of CE in probing the interaction of proteins with diverse particles. The separation power of CE should also facilitate studies of the multicomponent interaction systems for investigating how adsorption onto nanoparticles could affect the protein-protein or protein-small molecule interactions.
Li
Ni
N
Department of Chemistry, University of California, Riverside, California 92521-0403, USA.
Zeng
Shang
S
He
Le
L
Zhong
Wenwan
W
D019075:Electrophoresis, Capillary / Q000379:methods*; D052203:Ferrosoferric Oxide / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D011485:Protein Binding; D012492:Salts / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D012492:Salts; D012710:Serum Albumin, Bovine; D006046:Gold; D052203:Ferrosoferric Oxide