Characterization of Biological Material Adsorption to the Surface of Nanoparticles without a Prior Separation Step: a Case Study of Glioblastoma-Targeting Peptide and Lipid Nanocapsules

Purpose

Current preclinical therapeutic strategies involving nanomedicine require increasingly sophisticated nanosystems and the characterization of the complexity of such nanoassemblies is becoming a major issue. Accurate characterization is often the factor that can accelerate the translational approaches of nanomedicines and their pharmaceutical development to reach the clinic faster. We conducted a case study involving the adsorption of the NFL-TBS.40–63 (NFL) peptide (derived from neurofilaments) to the surface of lipid nanocapsules (LNCs) (a combined nanosystem used to target glioblastoma cells) to develop an analytical approach combining the separation and the quantification in a single step, leading to the characterization of the proportion of free peptide and thus the proportion of peptide adsorbed to the lipid nanocapsule surface.

Methods

LNC suspensions, NFL peptide solution and LNC/NFL peptide mixtures were characterized using a Size-Exclusion Chromatography method (with a chromatographic apparatus). In addition, this method was compared to centrifugal-filtration devices, currently used in literature for this case study.

Results

Combining the steps for separation and characterization in one single sequence improved the accuracy and robustness of the data and led to reproducible results. Moreover the data deviation observed for the centrifugal-filtration devices demonstrated the limits for this increasingly used characterization approach, explained by the poor separation quality and highlighting the importance for the method optimization. The high potential of the technique was shown, proving that H-bond and/or electrostatic interactions mediate adsorption of the NFL peptide to the surface of LNCs.

Conclusions

Used only as a characterization tool, the process using chromatographic apparatus is less time and solvent consuming than classical Size-Exclusion Chromatography columns only used for separation. It could be a promising tool for the scientific community for characterizing the interactions of other combinations of nanosystems and active biological agents.

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Article information: Gazaille, C., Sicot, M., Akiki, M. et al. Characterization of Biological Material Adsorption to the Surface of Nanoparticles without a Prior Separation Step: a Case Study of Glioblastoma-Targeting Peptide and Lipid Nanocapsules. Pharm Res (2021). https://doi.org/10.1007/s11095-021-03034-8

Materials: Span® 80 (sorbitane monooleate) (Span), sodium cholesteryl sulfate (SChol), and didodecyldimethylammonium bromide (DDAB) were supplied by Sigma-Aldrich (Saint-Quentin-Fallavier, France). Kolliphor® HS15 (mixture of free polyethylene glycol 660 and polyethylene glycol 660 hydroxystearate) (Kol) was purchased from BASF (Ludwigshafen, Germany). Labrafac® WL 1349 (caprylic-capric acid triglycerides) (Lab) was provided by Gattefossé S.A. (Saint-Priest, France). 100 K Amicon® Ultra filters (filter cut-off = 100 kDa) were supplied by Merck (Darmstadt, Allemagne). NaCl was purchased from Prolabo (Fontenay-sous-bois, France). Deionized water was obtained from a Milli-Q plus® system (Millipore, Bilerica, USA). The biotinylated NFL-TBS.40–63 peptide (NFL) was synthesized by the PolyPeptide group (Strasbourg, France).