Efficiently scaled-up production of recombinant human elastin-like polypeptides using multiple optimization...

Area of Science:

• Biotechnology
• Biomaterials Engineering
• Recombinant Protein Production

Background:

• Elastin-like polypeptides (ELPs) are biocompatible biopolymers with unique inverse transition cycling (ITC) properties, making them suitable for biomedical uses.
• Existing methods for ELP production suffer from low yields and complex purification, hindering scaled-up manufacturing.
• A need exists for efficient and cost-effective industrial production processes for ELPs.

Purpose of the Study:

• To develop an efficient, low-cost industrial production process for recombinant human elastin-like polypeptide (rhELP).
• To optimize rhELP expression and purification for large-scale manufacturing.
• To evaluate the biological activity of the produced rhELP.

Main Methods:

• Engineered an *Escherichia coli* (*E. coli*) strain for high rhELP expression, optimizing conditions using response surface methodology.
• Employed high-cell-density fermentation in automated fermenters to achieve high yields.
• Developed an efficient purification strategy involving homogenization, flocculation, salting out, hollow fiber membrane (HFM) filtration, ion-exchange chromatography (IEC), and ultrafiltration.

Main Results:

• Achieved a rhELP yield exceeding 5.00 g/L in automated fermenters, meeting industrial production demands.
• Purified rhELP to 99.83% purity with an 80.40% recovery rate using the optimized multi-step process.
• Demonstrated that the produced rhELP is non-cytotoxic and promotes wound healing both *in vitro* and *in vivo*.

Conclusions:

• This study establishes a universal paradigm for the industrial production of ELPs and similar recombinant proteins.
• The developed process significantly advances the commercialization potential of promising ELPs for biomedical applications.
• The efficient and scalable production method overcomes previous limitations in ELP manufacturing.