Spotlight on high-yield VHH expression using UNLOCK PICHIA

UNLOCK PICHIA - Productivity through diversity

VALIDOGEN´s PAOX1 promoter technology for protein production in Pichia pastoris provides several advantages. Very high titers along with timely adjusted expression of helper proteins (some available exclusively through VALIDOGEN) as well as lower consumption or complete absence of methanol are features which can significantly contribute to enhanced productivity and hence substantial reduction in cost of goods for protein manufacturing.

CASE STUDY: VHH product yield improvement through timely adjusted expression of helper proteins

In VALIDOGEN´s labs, promoter variants are applied simultaneously in pools of expression constructs comprising the gene of interest and potentially also genes encoding for different auxiliary (helper) proteins, thereby generating a broad diversity of genomic arrangements in the host cells resulting in diverse expression profiles. In this setting, protein production in Pichia pastoris is boosted significantly. Following specialized protocols, thousands of clones can be screened in minimum time and most promising clones will be reliably identified already at microscale. Screening results are transferable into bioreactors making laborious intermediate steps for clone comparison obsolete.

Profit from an extended helper factors portfolio acting at multiple sites within the production host

In an experimental case study a set of helper factors acting either in the cytosol or the ER or facilitating translocation of nascent Proteins into the ER were co-expressed in order to investigate a potential enhancement of VHH-dimer expression. Results from microscale as well as bioreactor scale showed a 2.0 - 2.7-fold improvement of product yield. Combining all three helper factors in one expression construct led to a 3.5-fold enhancement of product yield as depicted in the graph below.