Optimizing Desolvation Conditions for Glutathione-Cross-Linked Bovine Serum Albumin Nanoparticles: Implication for Intravenous Drug Delivery

Protein-based nanocarriers, particularly albumin nanoparticles (NPs), offer numerous advantages when compared to other nanomaterials. These carriers are characterized by biocompatibility, biodegradability, reduced immunogenicity, and decreased cytotoxicity. Moreover, proteins possess an inherent ability to target tumor cells directly or indirectly. This study aims to investigate the impact of various organic solvents on the characteristics of synthesized bovine serum albumin NPs (BSA NPs). BSA NPs were produced using methanol, acetone, ethanol, dimethylsulfoxide (DMSO), and acetonitrile through the desolvation technique to achieve particles of acceptable size. Dynamic light scattering (DLS), blood compatibility assays, polyacrylamide gel electrophoresis (PAGE), and size exclusion chromatography (SEC) were employed to elucidate the properties of the generated NPs. The cytotoxicity of BSA NPs prepared under different conditions was assessed using Michigan Cancer Foundation - Mammary Adenocarcinoma - Breast Cancer 231 cells (MDA-MB-231 cells). The particle size of the synthesized NPs varied based on the organic solvent utilized, with the smallest size of 114.2 nm observed with methanol. Blood compatibility results indicated no abnormal interactions between BSA NPs and blood components. PAGE analysis revealed a strong band near 72 kDa for untreated BSA and BSA treated with all organic solvents. In SEC, the retention time of native albumin was 6.65 min, while the average retention times of the prepared BSA NPs ranged from 5.14 to 5.21 min, showing similarity to the native protein. Except for NPs produced with methanol and acetonitrile, cytotoxicity testing on MDA-MB-231 cells demonstrated no significant harmful effects at various concentrations (0-500 μg/mL). The choice of desolvating agent significantly influences the size of BSA NPs. Various factors, such as solvent characteristics like hydrogen bonds, polarity, dielectric constant, and functional groups, can affect the particle size and structure of BSA NPs. The compatibility of cross-linked BSA NPs with blood components suggests their potential for intravenous drug delivery applications.