Investigating and Optimizing Insulin Partitioning with Conjugated Au Nanoparticles in Aqueous Two-Phase Systems Using...

Area of Science:

• Biochemistry
• Nanotechnology
• Separation Science

Background:

• Insulin is a critical biopharmaceutical requiring efficient purification methods.
• Aqueous two-phase systems (ATPS) offer biocompatible separation but face limitations for industrial biomolecule purification.

Purpose of the Study:

• To investigate the impact of bioconjugating insulin to gold nanoparticles (AuNPs/insulin) on its partitioning behavior in ATPS.
• To optimize ATPS parameters for maximizing the partition coefficient of AuNPs/insulin.

Main Methods:

• Synthesis and characterization of insulin-conjugated gold nanoparticles (AuNPs/insulin).
• Utilized polyethylene glycol (PEG)-dextran ATPS with varying pH, polymer molecular weights and concentrations, and AuNPs/insulin dosage.
• Employed response surface methodology (RSM) with a D-optimal design for system modeling and optimization.

Main Results:

• Insulin retained full activity after conjugation to gold nanoparticles.
• Optimized conditions (pH 8, 21% PEG 4000, 5% Dextran 100,000, 100 IU AuNPs/insulin) yielded a partition coefficient of 192.96 for AuNPs/insulin.
• Achieved partition coefficient for AuNPs/insulin was significantly higher than that of free insulin.

Conclusions:

• Bioconjugation of insulin to gold nanoparticles dramatically improves its separation efficiency in ATPS.
• This strategy overcomes limitations of ATPS for industrial-scale purification of biomolecules and biopharmaceuticals.
• The developed method holds potential for advancing biopharmaceutical manufacturing processes.