Process development of a FGF21 protein-antibody conjugate

Area of Science:

• Biopharmaceutical manufacturing
• Protein engineering
• Therapeutic protein development

Background:

• Fibroblast Growth Factor 21 (FGF21) protein-antibody conjugates, like CVX-343, are novel therapeutics for metabolic diseases.
• Complex bioconjugate manufacturing requires optimized processes for scalability and viability.
• Existing CVX-343 manufacturing had challenges requiring improvements for Phase 3 and commercial production.

Purpose of the Study:

• To develop a scalable and viable manufacturing process for the FGF21-protein conjugate CVX-343.
• To enhance the stability of intermediates and minimize dimerization and hydrolysis.
• To optimize reaction conditions for improved efficiency and product quality.

Main Methods:

• Eliminated tris(2-carboxyethyl)phosphine (TCEP) to prevent disulfide-mediated FGF21 dimerization.
• Formulated and activated FGF21 at acidic pH to minimize dimerization and linker hydrolysis.
• Utilized an activation use test for FGF21 pooling to reduce misfolds and dimers.

Main Results:

• Reduced FGF21 consumption by 36% and linker consumption by 55%.
• Decreased overall process time from approximately 5 days to 3 days.
• Improved product distribution of the desired bifunctionalized conjugate from ~60% to ~75% in crude mixture and ~80% to ~85% in final drug substance.

Conclusions:

• An optimized, scalable manufacturing process for CVX-343 was successfully established.
• The new process enhances efficiency, reduces material consumption, and improves product quality.
• This optimized process supports the advancement of CVX-343 for commercial manufacturing.