Engineered BSA Nanoparticles: Synthesis, Drug Loading, and Advanced Characterization

Abstract Bovine serum albumin (BSA) nanoparticles have attracted a lot of interest as biocompatible and biodegradable carriers for a range of pharmacological and biological uses. BSA nanoparticles have several advantages over other types of nanoparticles, including their ability to increase the stability and solubility of encapsulated drugs, their non-toxicity, and their ease of surface modification. Cancer treatment, immunological modulation, enzyme immobilization, controlled release systems, bioimaging, and theranostics are some of its potential applications. This protocol offers a detailed and accessible methodology for the synthesis, drug encapsulation, and characterization of albumin nanoparticles with an emphasis on reproducibility and adaptability. The synthesis uses the desolvation process and crosslinking with compounds glutaraldehyde for stability. The crosslinking ratio, pH, and BSA content are important factors that are adjusted to control size, surface charge, and dispersity. The methods used for characterization are described in detail, including dynamic light scattering (DLS) for particle size and zeta potential, transmission and scanning electron microscopy (TEM/SEM) for morphology, Fourier-transform infrared spectroscopy (FTIR), and nanoparticle tracking analysis (NTA) for stability assessment. The stability of the nanoparticles was evaluated under physiologically relevant ionic and pH conditions by dispersing them in phosphate-buffered saline (PBS), providing insight into their colloidal behavior in a simulated physiological environment. This technique facilitates the design of functionalized BSA nanoparticles for certain biomedical and therapeutic applications by acting as a fundamental reference for researchers. This work promotes innovation in nanoparticle-based technology and advances the field by standardizing preparation and characterization techniques.