Quantification of BSA-loaded chitosan/oligonucleotide nanoparticles using reverse-phase high-performance liquid...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Materials Science

Background:

• Therapeutic proteins require careful delivery due to gastrointestinal instability.
• Polymeric nanoparticles, microparticles, and liposomes are promising nanocarriers for protein encapsulation.
• Accurate quantification of proteins is essential for evaluating drug delivery systems, but excipients can interfere with analytical methods.

Purpose of the Study:

• To investigate the impact of common polymers on bovine serum albumin (BSA) quantification using RP-HPLC.
• To validate a RP-HPLC method for analyzing protein release from nanoparticle formulations.
• To characterize the in vitro drug release profile of a novel BSA-loaded nanoparticulate system.

Main Methods:

• Evaluated the effect of nine common polymers on BSA quantification via RP-HPLC.
• Developed and validated a RP-HPLC method for protein separation and quantification.
• Formulated BSA-loaded nanoparticles using chitosan and oligonucleotide (Dz13Scr) via complex coacervation.
• Characterized nanoparticle size, polydispersity index, and surface charge.
• Assessed in vitro drug release in simulated gastric and intestinal fluids over 12 hours using the validated RP-HPLC method.

Main Results:

• The RP-HPLC method demonstrated high accuracy, selectivity, and short analytical time for BSA quantification.
• The method successfully separated and quantified BSA, insulin, and pigment epithelium-derived factor (PEDF).
• Novel BSA-loaded nanoparticles exhibited favorable characteristics: 337.87 nm size, 0.338 PDI, and +10.23 mV charge.
• The validated RP-HPLC method effectively quantified the in vitro release of BSA from the nanoparticles.

Conclusions:

• RP-HPLC is a suitable analytical technique for quantifying therapeutic proteins in the presence of polymeric excipients.
• The developed RP-HPLC method is effective for characterizing drug release from nanocarrier systems.
• The novel chitosan-oligonucleotide nanoparticles show potential as a delivery system for therapeutic proteins, with their release profile accurately determined by RP-HPLC.