Premium vector technology driving improved productivity and quality

In mammalian cell lines
Improve the expression of recombinant proteins in all mammalian platforms

Additional applications
Discover even more novel applications that can benefit from our technology
Improve the expression of your recombinant proteins in mammalian platforms
2G UNic™: premium vector technology for increased protein production levels
A combination of proprietary genetic elements in one vector, together driving higher protein production levels

More mRNA per integrated DNA copy
Increased transcription levels due to:
- Double promoter: two promoters are combined in a novel way to increase transcription
- Epigenetic stabilization: reduced epigenetic down-regulation allows stable high-level expression
More protein per mRNA molecule
- Improved translation efficiency by optimized ribosome affinity
- Optimized mRNA transport by use of a synthetic intron
- Optimized mRNA stability
- Superior performance: multi-fold increase in recombinant protein productivity over the entire range of expression levels:
- Low level productivity (< 1 g/l) to commercially viable levels
- Medium to high levels, e.g., from 2 to 5 g/l
- High end levels, e.g., from 5 to 8 g/l
- Broad applicability:
- Across all relevant mammalian cell lines and expression systems
- For different types of molecules
- For random and targeted integration; multi- and single copy
- Easy process integration: upgrade current processes with minimal changes to systems, workflow or product quality
- Reduce screening effort: fewer resources required to identify high-producing clones
- Compatible and synergistic: combine with other expression-enhancing technologies to further boost productivity, e.g., transposon or chromatin modulation technology
- Significant savings: potentially save millions of euros per year, per product (based on typical volumes)
- Further growth opportunities: can be applied to other in-demand areas, e.g., viral vector and vaccine production, mRNA therapeutics, production in yeast and other fungal cells, etc.
- Strong IP position: Fully owned IP with four independent patent families in the US and Europe
Suitable for a broad range of proteins, cell types, and selection markers, including:
Interested in trying 2G UNic™ for your application?
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- Easy and efficient vector cloning: one-step construction
- Compatible with GoldenGate assembly
- Mono- and bicistronic vector formats available
- Straightforward preparation of transfection-ready linear DNA by Shredder technology (patent pending)
Conclusion: Superior titer obtained using 2G UNic™
Protein: Bispecific antibody
Cell line: CHO GS-/- (HD-BIOP3)
Approach: 2G UNic™ vector
Results: 3-fold increased titers with 2G UNic™ vectors
ST6 sialylation technology: a glyco-engineering approach for improved protein quality
Achieve human-type sialylation of biotherapeutic proteins in CHO cells for improved efficacy and reduced immunogenicity

- High yield & superior protein quality combined
- Easy process integration: apply by simple co-transfection
- CHO cell line compatible, including GS and DHFR knockout lines
- Replaces human cell platforms: sialylation levels similar to serum IgGs and IVIg under standard culture conditions
- Versatile sialyl transferase vector:
- Combined with 2G UNic™ vectors provides high biotherapeutic titers with superior product quality in any CHO system
- Licensed separately for transfection into an existing cell line
- Analysis and optimization services and consultancy available
A chimeric ST6 minigene in an expression vector with selectable marker
- ST6 activity is absent in wild-type CHO cells and expressed proteins are poorly sialylated
- One-step approach for human-type 6-sialylation by stable expression of novel modified ST6
- Efficient human-type 6-sialylation of N-glycans in the Fc region of IgGs and Fc-fusion proteins in CHO cells
Conclusion: Human serum-like sialylation of mAb achieved by co-transfection with novel ST6 Protein: IgG1 Cell line: CHOZN; bulk pools Approach: Co-transfection of 2G UNic™ vectors for mAb and ST6 Results:
- Increased sialylation level to ≈15% with ST6, similar to intravenous immunoglobulins (IVIg)
- ST6 has a minimal impact on IgG1 expression levels
- Sialylation levels >75% can be obtained by clone selection and feed optimization while maintaining high expression levels

Improve expression levels in yeast platforms
2G-P UNic™: premium vector technology for increased protein production levels in yeast
Premium expression vectors for fungal and yeast protein production platforms

2G-P UNic™ technology has been optimized and validated for reliable application
- Severalfold higher higher protein production levels
- Scalable: yields stable, or further improved, in fed batch bioreactor runs
- Reproducible: production increased in >90% of proteins tested
- Independent of promoter type: >2-fold yield increase achieved with constitutive and inducible promoters
- Pichia compatible: multifold titer enhancement achieved in Pichia
- Broadly applicable: applications beyond biopharmaceuticals include industrial enzymes, food, feed
- Validated by external parties
2G-P UNic™ is a combination of novel yeast-specific genetic elements which together improve recombinant protein expression in yeast cells
In analogy to ProteoNic's expression vectors for mammalian platforms, 2G-P UNic™ increases transcription and translation rates, resulting in:
- More mRNA per integrated DNA copy
- More protein per mRNA molecule
Are you interested in trying 2G-P UNic™ for your application?
Effect of 2G-P UNic™ on production of a biopharmaceutical protein in yeast (Pichia pastoris)
Approach: Strain selection with copy-number amplification; comparison of best reference strain with best 2G-P UNic™ strain
Results: 6-fold yield increase in 30 L bioreactor
We are continuously developing new applications for our protein-boosting technology
Additional applications of ProteoNic's technology

2G UNic™ is applicable to a wide range of other applications:
- Viral vectors driving improved viral particle production for gene therapy
- Improved gene therapy efficacy
- Transient protein expression
- Baculovirus-based production in insect cells
- mRNA-based therapeutic development
- Vaccine production
- Various non-human or non-medical applications
Interested in trying 2G UNic™ for your application?