VALIDOGEN White Paper: Boosted expression of VHH antibodies through application of UNLOCK PICHIA tools

white-paperVALIDOGEN GmbH
November 24th 2020

Protein expression specialist VALIDOGEN has published a White Paper that shows how expression of a range of different VHH antibody formats can be increased through application of different components of its proprietary UNLOCK PICHIA toolbox.

The studies are significant because VHHs, derived from camelid species, the so-called “third generation” of antibodies and also known as sdAbs or Nanobodies®, exhibit characteristics that make them superior to mAbs and antibody fragments (Fab and scFv) for many applications. These advantages include high specificities and affinities in antigen binding, along with small size, high solubility, thermal stability, design flexibility and modularity, good tissue penetration in vivo, and expression in microbial systems. In consequence, VHHs are currently being developed for an increasing number of different applications including pharmaceutical use as APIs, in diagnostics as well as for diverse applications in other industries.


The case study demonstrates, among others, the benefits of applying a higher number of PAOX1 promoter variants from VALIDOGEN’s UNLOCK PICHIA toolbox in a ‘pool approach’ which typically leads to a significant productivity boost, at the same time enabling more rapid generation and identification of high-level production strains. The targeted application of VALIDOGEN’s full toolbox by identification of optimum combinations with different elements such as strain background, helper factors, secretion signals or production regimes further boosts productivity.

The results of the studies confirm the advantages of VALIDOGEN´s broad UNLOCK PICHIA platform featuring numerous tools and different expression strategies for the enhancement of production yields and product quality, delivering key advantages:

  • Boosted expression levels for monovalent- and multivalent VHH antibodies: up to 16.5 g/l
  • Secreted products providing an appealing starting point for protein purification
  • Complete absence of methanol through VALIDOGEN´s methanol-free promoters
  • Stable, robust and scalable processes for commercial manufacturing.

VALIDOGEN White Paper Boosted expression of VHH antibodies through application of UNLOCK PICHIA tools


Click on UNLOCK PICHIA for further information on Pichia pastoris production toolbox