At NIZO we have:
• a closed and well de-aerated robotic screening setup available for designing and testing media. Sterilization and de-aeration of complex media in UHT systems ensures proper growth of bacteria, selected from these niches and maintain maximal nutritional value whilst minimizing Maillard-reactions.
• A high & medium throughput probiotic optimization pipeline (96 well plate, 0.5 L and 2 L vats) for fully anaerobic and efficient screening in a single assay. The full production process including growth, concentration, lyophilization and shelf-life stability testing is downscaled to We screen for medium, for interaction effects of different medium components pH’s and cryoprotectants, and for the optimal growth phase for harvesting.
• Medium scale DSP optimization at the 2 liter scale allows for testing the influence of downstream processing using industrial scalable technologies as cross-flow filtration and centrifugation for concentration prior to lyophilization of the cells and shelflife stability testing.

‘Next generation’ probiotics are originating from niches like the (human) gut, skin or oral cavity. These live biotherapeutic products often prove very difficult to culture as many of them have never been used in industrial applications and a large number of them are strict anaerobes.

To optimize fermentation yield, cell robustness and shelf life stability for our customers’ probiotic product, NIZO can test thousands of different combinations of up- and downstream production processing conditions. This allows identification of optimal combinations of:
1. growth conditions: strains and media/substrate
2. freeze dry conditions: harvest moments and choice of cryoprotectants
3. shelf life stability

For these tests we developed the high throughput Probiotics Optimization Pipeline that allows full screening of important properties (such as growth characteristics, enzyme activities and metabolites) in thousands of different culture conditions, combining our extensive knowledge of engineering of lactic acid bacteria (through natural as well as DNA modifying techniques) with our microplatforms and genomics and proteomics platforms. The probiotics pipeline is deployed for traditional probiotic strains as well as for strict anaerobic species, such as gut-derived bacteria and skin-derived bacteria.

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Benefits of the probiotics pipeline are:
1. Risk Reduction For Your Probiotic Production Process
The Probiotic Optimization Pipeline allows testing of a large matrix of thousands of media, cryopreservative, harvesting phase and strain combinations. The best performing process conditions may be pre-selected for further analysis at the 0.5 liter scale. This allows for a strongly de-risked research phase at a scale that allows easy up-scaling without having to test a large set of processing conditions in order to identify the right processing conditions for your process

2. Smaller Scale, Better Price
Performing thousands of CFU analyses using standard technologies would require many resources (manpower, materials) and would thus be cost prohibitive. NIZO has been able to perform high throughput CFU analyses of both the aerobic and anaerobic fermentations thereby allowing for testing of many DSP conditions and shelf life stability tests from samples derived from any scale of anaerobic or aerobic fermentation

In our pilot plant we offer the following upscaling and production possibilities to our customers:
• Upscaling approach aligned with final GMP requirements (contacts with CMO are established)
• 40L, 650L, 4500L fermentation capacity in food grade environment
• Flexible DSP options (centrifugation, cross-flow filtration) and formulations (liquid, powder, encapsulated)
• (Accelerated) shelf life testing
• Many types of packaging are supported
• Quality checks for release of products according to your preferred guidelines
• Tech transfer of process to GMP producer

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