Conjugation of hydroxyapatite nanocrystals with human immunoglobulin G for nanomedical applications

• Published in: Colloids and surfaces, B, Biointerfaces B, Biointerfaces, 2012-02, Vol.90, p.1-7
• Main Authors: Iafisco, Michele, Varoni, Elena, Di Foggia, Michele, Pietronave, Stefano, Fini, Milena, Roveri, Norberto, Rimondini, Lia, Prat, Maria
• Format: Article
• Language: English
• Subjects: Adsorption; Antibodies; Antibody; antigens; Antineoplastic Agents - chemistry; Antineoplastic Agents - immunology; Antineoplastic Agents - metabolism; Binding Sites; biomimetics; Blotting, Western; calorimetry; colloids; dose response; drug carriers; Drug Carriers - chemistry; Drug Carriers - metabolism; drugs; Durapatite - chemistry; Durapatite - immunology; Enzyme-Linked Immunosorbent Assay; Functionalization; Humans; Hydroxyapatite; Immunoconjugates - chemistry; Immunoconjugates - immunology; Immunoconjugates - metabolism; immunoglobulin G; Immunoglobulin G - chemistry; Immunoglobulin G - immunology; Immunoglobulin G - metabolism; Mathematical models; Molecular Targeted Therapy - methods; Nanocrystals; Nanomaterials; Nanomedicine; Nanomedicine - methods; Nanoparticles; Nanoparticles - chemistry; Nanostructure; neoplasms; Neoplasms - immunology; Neoplasms - therapy; Particle Size; Protein Binding; Protein Structure, Secondary; sorption isotherms; spectroscopy; Spectrum Analysis; Static Electricity; Surface chemistry; Surface Properties; Western blotting
• ISSN: 0927-7765; 1873-4367
• DOI: 10.1016/j.colsurfb.2011.09.033

[Display omitted] ► Biomimetic hydroxyapatite nanocrystals synthesized and conjugated with human IgG. ► IgG adsorption is dose-dependent, saturable, stable, with a capability of 2.3 mg/m 2. ► Binding is mainly due to electrostatic interactions, with no IgG structure modification. ► Preferential IgG end-on orientation, probably exposing Fab domains at the surface. ► Hydroxyapatite nanocrystals could be suitable carriers for targeted drug delivery. Inorganic nanosized drug carriers are a promising field in nanomedicine applied to cancer. Their conjugation with antibodies combines the properties of the nanoparticles themselves with the specific and selective recognition ability of the antibodies to antigens. Biomimetic carbonate–hydroxyapatite (HA) nanoparticles were synthesized and fully characterized; human IgGs, used as model antibodies, were coupled to these nanocrystals. The maximum loading amount, the interaction modelling, the preferential orientation and the secondary structure modifications were evaluated using theoretical models (Langmuir, Freundlich and Langmuir–Freundlich) spectroscopic (UV–Vis, Raman), calorimetric (TGA), and immunochemical techniques (ELISA, Western Blot). HA nanoparticles of about 30 nm adsorbed human IgGs, in a dose-dependent, saturable and stable manner with micromolar affinity and adsorption capability around 2.3 mg/m 2. Adsorption isotherm could be described by Langmuir–Freundlich model, and was due to both energetically homogeneous and heterogeneous binding sites on HA surface, mainly of electrostatic nature. Binding did not induce secondary structure modification of IgGs. A preferential IgG end-on orientation with the involvement of IgG Fc moiety in the adsorption seems most probable due to the steric hindrance of their Fab domains. Biomimetic HA nanocrystals are suitable substrates to produce nanoparticles which can be functionalized with antibodies for efficient targeted drug delivery to tumours.