Modeling, simulation, and employing dilution-dialysis microfluidic chip (DDMC) for heightening proteins refolding...

Area of Science:

• Biochemical Engineering
• Microfluidics
• Protein Chemistry

Background:

• Microfluidic systems are crucial for biochemical and biomedical applications, requiring systematic design through mathematical simulations.
• Protein refolding from inclusion bodies is vital for industrial recombinant protein production, often relying on denaturant dilution or dialysis.
• Current methods face challenges in efficiently suppressing protein aggregation during refolding.

Purpose of the Study:

• To introduce and model a compact, cost-effective on-chip refolding method using microfluidics.
• To develop an innovative dilution-dialysis microfluidic chip (DDMC) for improved protein refolding.
• To validate the DDMC's effectiveness for controlled urea reduction and enhanced protein refolding.

Main Methods:

• Developing a dilution-dialysis microfluidic chip (DDMC) integrating gradual urea reduction and stepwise dialysis.
• Modeling the DDMC operation in two dimensions using COMSOL Multiphysics.
• Conducting experimental validation using Hen egg white lysozyme (HEWL) as a model protein.

Main Results:

• The DDMC design was simulated and found suitable for controlled urea decrease.
• Experimental results demonstrated significantly improved refolding efficiency using the DDMC.
• Refolding efficiency for HEWL increased from 42.6% with simple dilution to 93.7% with DDMC.

Conclusions:

• The DDMC offers a promising, efficient, and cost-effective approach for on-chip protein refolding.
• The microfluidic system effectively controls urea concentration, enhancing native protein recovery.
• This technology has significant potential for industrial applications in recombinant protein production.