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VALIDOGEN protein expression services for effective recombinant protein production
VALIDOGEN GmbH (formerly VTU Technology) offers services based on a world-leading versatile and effective toolbox and production platform for Pichia pastoris recombinant protein expression and production known as UNLOCK PICHIA. VALIDOGEN’s UNLOCK PICHIA technology platform comprises a suite of expression tools for the reliable & rapid generation and straightforward identification of effective industrial protein production strains without compromising development timelines.
The yield-enhancing and industry proven yeast protein expression platform combines proprietary AOX1 promoter libraries and expression-enhancing helper factors, diverse secretion signals, various optimized platform strains, elaborated cloning and transformation protocols and a high-throughput microscale screening and cultivation regime combined with excellent know-how in fermentation and downstream process development for maximization of product yield and overall process performance. The platform enables high-titer production with record breaking productivities of up to 35 g/L of secreted protein.
VALIDOGEN’s suite of UNLOCK PICHIA protein development and production services
VALIDOGEN develops customized high-performance Pichia pastoris production strains at competitive timelines and offers straightforward bioreactor cultivation & protein purification process development and optimization services paired with technology transfer and/or lab-scale protein production.
Protein expression services:
- Protein expression studies: Rapid assessment of expression potential, using streamlined and time saving feasibility studies
- Comprehensive strain development: Targeted and time-saving generation of high-productivity strains already considering economic target values and regulatory requirements applying VALIDOGEN’s broad UNLOCK PICHIA toolbox
- Bioprocess development: Straightforward fermentation development, optimization and characterization services
- Protein purification and DSP development: Protein purification and product characterization, broad toolbox of established analytical methods
- Cell banking: Preparation and characterization of research cell banks
- Enzyme engineering: High-throughput engineering and screening of enzyme variants
Protein expression studies and comprehensive strain development
VALIDOGEN offers tailored protein expression studies as the starting phase for any customer development program that requires initial determination of expression potential and rapid production of small scale protein samples. These studies are particularly useful where expressibility needs to be assessed at early stage and where expression in other commercially available protein expression systems has proved problematic and where client needs scale up to industrial protein production.
A lean feasibility study will start from an optimized synthetic gene sequence to elucidate the expression potential of recombinant protein candidates in minimum time applying a subset of VALIDOGEN’s AOX1 promoter library, with protein synthesis results available within a few weeks. This gives the customer fast and reliable data to support a go/no-go decision.
VALIDOGEN’s custom protein expression studies and strain development program are based on both methanol-induced or methanol-free processes. Protein expression constructs carrying UNLOCK PICHIA promoter variants, different secretion signal peptides, the gene of interest and potentially also genes encoding for different auxiliary (helper) proteins can be transformed into selected Pichia platform strains, generating a broad diversity of genetic arrangements in the host cells for diverse expression profiles and significantly boosting protein production in Pichia pastoris.
Following specifically optimized protocols, thousands of clones can be screened in minimum time with most promising clones reliably identified at microscale. Screening results are transferable into bioreactors, rendering obsolete those laborious intermediate steps that have been conventionally used for clone comparison.
Following positive assessment, VALIDOGEN can then provide yield optimization by applying the full set of its proprietary comprehensive protein production strain development tools.
UNLOCK PICHIA advantages
VALIDOGEN’s protein expression services harness all the UNLOCK PICHIA toolbox elements to generate a large number of different genetic constellations in thousands of strains for fine-tuning of expression, unlocking the power of Pichia pastoris for successful production of fully active, high-quality proteins.
In summary, UNLOCK PICHIA provides a very broad range Pichia-based platform considering all aspects of manufacturability from targeted debottlenecking for yield maximization, to product quality (including glycosylation issues) and regulatory requirements (e.g. antibiotic-free clone selection) as well as process safety (such as methanol-free production) and process economy are considered and integrated at the earliest possible stage during strain and process development. Hence, effective industrial protein production strain generation and production process development is assured paving the way for economically viable manufacturing.
- Boosted expression levels
- Short cultivation times
- Secreted products providing a beneficial starting point for protein purification
- Complete absence of methanol through VALIDOGEN´s methanol-free promoters
- Stable, robust and scalable processes for commercial manufacturing.