Exceptional Productivity of 31 g/L for a bivalent VHH with VALIDOGEN’s UNLOCK PICHIA? Platform

CASE STUDY III: Methanol-induced bivalent VHH production

Applying our PAOX1-driven, methanol-induced UNLOCK PICHIA? process we achieved impressive protein expression levels, reaching 31 g/L for a bivalent VHH in Pichia pastoris.

Key Achievements

ENHANCED STRAIN ENGINEERING LEVERAGING UNLOCK PICHIA? TECHNOLOGY: By harnessing our UNLOCK PICHIA? toolbox, we developed a high-performance Pichia pastoris strain capable of secreting 21 g/L of bivalent VHH applying one of our generic standard protocols.

ADVANCED PROCESS OPTIMIZATION: Through sophisticated process development and optimization strategies we achieved a ~50% increase in product titer, boosting final yields to an impressive 31 g/L in just 111 hours cultivation.

Productivity Highlights

SPECIFIC PRODUCTIVITY: Improved cultivation conditions through process optimization and scale-up significantly boosted cell performance, resulting in a remarkable ~80% increase in specific productivity.

Case Study III: Development of a methanol-induced Pichia protein production process for maximization of process performance

We previously published the findings of Case Studies I and II, showcasing the development of methanol-free production processes for the efficient production of a bivalent VHH. Through process optimization, we achieved product yields of 12 g/L in just 64 hours and 19 g/L within VALIDOGEN's standard cultivation time of 111 hours.

These processes demonstrated exceptionally high space-time yields (STY), providing a safe and efficient alternative for protein production processes where methanol needs to be avoided.

Case study III aimed to develop a high-performance methanol-inducible production process for the expression of the same molecule, a bivalent VHH.

Using a VALIDOGEN generic standard protocol 21 g/L of bivalent VHH was secreted in 1 L bioreactors. In an initial development phase existing VALIDOGEN optimization protocols were applied to this strain in small-scale trials. This resulted in an increase of yield to 24 g/L, and a notable improvement in specific productivity.

By subsequently applying a targeted process development and optimization strategy at the 10 L bioreactor scale we achieved an impressive titer of 31 g/L, alongside a remarkable ~80% improvement in specific productivity compared to the standard methanol process. A customized methanol feeding strategy was developed specifically for this strain, enabling both a successful scale-up and further enhancement of productivity.

These results demonstrate the exceptional potential of VALIDOGEN’s UNLOCK PICHIA? platform and Pichia expert know-how to develop high-performance protein production processes for commercial application.

Our platform provides extensive customization options, including both methanol-induced and methanol-free expression strategies. This flexibility, combined with our expertise in Pichia process development & optimization, enables us to develop robust, scalable, and economically viable processes that significantly reduce the cost of goods for protein manufacturing.

Contact us to learn more about VALIDOGEN's strain development & process development programs.

Iskandar Dib, Principal Scientist and BD Manager, [email protected]

Rosie Maddock, BD Manager, [email protected]