Methanol-induced protein expression in Pichia pastoris

Efficient and industry-proven Pichia expression system for maximization of protein yields

Methanol-induced protein production with UNLOCK PICHIA® – High titers, proven performance


At VALIDOGEN, we leverage our proprietary high-performance AOX1 promoter variants - key elements of our UNLOCK PICHIA® toolbox - with additional expression-enhancing components to boost protein production. Methanol-induced expression remains one of the most powerful strategies for many recombinant proteins, with VALIDOGEN’s system achieving high titers across a broad protein spectrum (peak productivities of up to 35 g/L of secreted protein).

With over 15 years of experience in developing industrial-scale production processes for both GMP and non-GMP customers across diverse sectors, VALIDOGEN has established advanced fermentation strategies that reliably overcome key challenges of methanol-based processes. The outcome: robust, scalable, and highly productive fermentation processes.

For projects where maximizing titer is essential and methanol (MeOH) handling is operationally viable, this system offers a compelling, efficient, and industry-proven solution.

If MeOH-free protein production is preferred - whether due to safety considerations, operational constraints, or regulatory requirements - we offer a proven, industry-leading alternative to methanol-induced systems, delivering performance that surpasses many other commercial platforms.
 

Benefits of VALIDOGEN's methanol-induced Pichia system

  • UNLOCK PICHIA® methanol-induced processes leverage engineered AOX1 promoters for high-performance protein expression
  • Methanol-inducible protein expression allows strong, tightly-controlled protein production (record productivity levels of 35 g/L of secreted protein)
  • Optimized standard cultivation protocols and strategies for maximization of product titers and specific productivity
  • Our long-standing know-how in handling methanol-based expression enables robust and scalable bioprocess solutions

     

Case Studies

Case studies highlight the powerful capabilities of our methanol-induced Pichia system for high-yield protein expression.

Methanol-induced bivalent VHH expression in Pichia pastoris

We recently published a three-part case study on the expression of bivalent VHH. While Case Study 1 & 2 focused on the development of methanol-free process optimization strategies, Case Study 3 highlighted the development of methanol-induced Pichia strains and protein production processes.
Using a VALIDOGEN standard protocol, 21 g/L of bivalent VHH was secreted in 1 L bioreactors. During the initial development phase, existing VALIDOGEN optimization protocols were applied to this strain in small-scale trials, resulting in a yield increase to 24 g/L and a notable improvement in specific productivity.

Further bespoke process development and optimization at the 10 L bioreactor scale led to an increased titer of 31 g/L, with a remarkable ~80% improvement in specific productivity compared to the standard methanol process. A customized methanol feeding strategy was developed specifically for this strain, facilitating both successful scale-up and further enhancement of productivity.

Read the full case study here.