From idea to first protein to product

Bioneer addresses the need for recombinant proteins and is typically involved in the production of the initial amount of protein for the discovery and preclinical phase. Bioneer exploits a variety of microbial and mammalian expression systems in order to fulfil this need. If a protein is successfully produced, we can then optimize the productivity by genetic and/or physiological methods. Our downstream processing unit enables purification at analytical scale and at gram scale. We have a proven track record in transferring processes developed at Bioneer to CMO’s. Lastly, we carry out all types of pre-formulation services including protein stability tests in collaboration with partners experienced in turning a protein into a product.

For further information, please contact Head of Department Søren Madsen by phone +45 45160444 or email.

Feasibility Studies

Bioneer offers expertise in recombinant protein production using bacterial and mammalian expression systems such as Escherichia coli, Lactococcus lactis, Chinese hamster ovary cells (CHO) and Human embryonic kidney cells 293 (HEK293).

In collaboration with third parties, we design and produce synthetic and codon optimized genes for recombinant expression in the desired host organism. We perform gene cloning into selected expression vectors and transform bacterial strains and transfect mammalian cell lines.

We generate research cell banks and perform QA characterization, e.g. by DNA sequencing. Cell banks can subsequently be transferred to CMOs for cell banking according to cGMP guidelines.

We optimize recombinant protein expression by varying growth media, temperatures and/or other expression-inducing conditions.

We analyze recombinant protein production by cellular fractionation of e. g cytoplasm or periplasm from coli. Yield and quality can be analyzed by SDS-PAGE, Western blotting or activity assays.

Upstream Development

Our bacterial fermentation facility hosts six Applikon bioreactors for development and process optimization in small scale. A 20 L stainless steel bioreactor  is available for a larger scale. Mammalian cells are cultivated in shaker flasks of different sizes.

We develop processes for batch and fed-batch (high-cell-density) fermentations. High protein production is achieved through optimization of physical and chemical induction conditions such as e. g IPTG concentration and temperature.

We perform small scale mammalian cell cultivation in 3 L flasks, and can run multiple cultivation flasks at a time.

Process optimization is based on the physiological needs of the microorganisms and of the mammalian cell lines.

In close collaboration with our clients, we prepare a tech transfer package securing smooth transfer to CMO responsible for cGMP manufacturing.

Downstream Development

Our protein purification facility is equipped with several Akta purification stations allowing development of various chromatographic purification strategies. The facility is also equipped with a high pressure homogenizer for extraction of intracellular proteins.

We have equipment for clarification of fermentation broth by centrifugation and tangential flow filtration. For extraction of intracellular proteins a high pressure homogenizer is available. Periplasmic proteins are extracted from E. coli using an osmotic shock procedure.

Recombinant proteins accumulating as inclusion bodies are solubilized and refolded by dilution or dialysis into appropriate buffer solutions.

We offer a variety of column-based purifications methods based on affinity, ion-exchange, hydrophobic interaction, mixed mode and gel filtration chromatography. Purification in gram scale is possible.

We can characterize the final recombinant protein with respect to quantity and quality using techniques and methods like:

  • Electrophoresis (SDS-PAGE, native PAGE, IEF)
  • Bradford and BCA protein quantification
  • UV spectrophotometry
  • Western blotting
  • HPLC (SEC, IEX and reversed phase chromatography)
  • Enzyme assays
  • Endotoxin measurement
  • HCP analysis
  • Circular dichroism
  • Dynamic light scattering

Protein Pre-Formulation

Bioneer and partners design and perform the critical experiments, which are necessary for creating a stable protein formulated according to regulatory requirements.

In collaboration with private partners, who have a proven track record in bringing a protein/a biologics through the pre-clinical and clinical phases, we develop a strategy for how a given protein can be stabililized by specific formulations to fulfill stability criteria necessesary for later regulatory approval.

Furthermore, we design pre-formulation experiments including research-based stability analysis in order to enable a smoother and also cost-effective transition of a protein from the research phase to the clinical phases.

Create new product opportunities

We offer access to essential tools, technology and unique biological resources within cell and molecular biology.

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