Probiotics: screening, optimization and production

At NIZO, we understand that selecting the right probiotic strain with the desired functionality and health benefits, as well as ensuring its manufacturability, is crucial for the success of your product. We offer a wide range of screening options to characterize the physiological properties of strains and substantiate their health benefits. With our advanced facilities, we can also produce material for clinical trials in our food-grade pilot plant and develop processes that can be transferred to GMP manufacturers. In addition to working with standard species, we have extensive experience with strict anaerobic gut commensals. Here are some examples of our activities in this field.

Discovery of microbiome modulators (in silico/HTS)

At NIZO, we begin the discovery process with state-of-the-art bioinformatics and machine learning techniques to identify enzymes, functions, and ultimately strains of interest based on genomics. With this knowledge, we can either regulate or monitor microbiome functions in vitro systems that are relevant. Our MicroColon and MicroSkin screening platforms enable the high-throughput assessment of new ingredients and probiotics for their effectiveness in the gastrointestinal tract and skin, respectively. Both models ensure a reliable translation of microbiome research for subsequent preclinical studies.

Medium development

We have developed a Probiotic Optimization Pipeline that enables us to test thousands of media and strain combinations to identify the best performing combinations. Our pipeline includes testing with LABs, strict anaerobic gut bacteria (such as Akkermansia, Clostridium, and Faecalibacterium) and skin commensals (such as Cutibacterium and Staphylococcus) to ensure growth and manufacturability. This process allows us to design animal-free media while minimizing medium complexity. The top performing combinations are then further analyzed at varying scales (0.5, 2, 30, and 600 liters) to optimize the medium/strain/process combination and maximize yield or functional activity of interest.

USP/DSP/Formulation

The survival of probiotic cells and their delivery to the consumer in a viable form largely depends on Upstream Processing (USP), Downstream Processing (DSP), and the final formulation. In our Probiotic Optimization Pipeline, we investigate different cryo- and lyoprotectants and their impact on cell survival. At NIZO, we have extensive expertise in freezing, freeze-drying, and spray-drying of microbes, as well as in the selection and optimization of preservation techniques. Our rational approach to process optimization ensures optimal cell viability and survival, which are crucial for the success of your probiotic product.

Upscaling 0.5, 40, 600, 4500 liter scale

Our high throughput platform which can be run in a strictly anaerobic environment typically leads to conditions to be further upscaled to produce probiotic strains. Optimization at the 0.5 litre scale allows for testing the influence of downstream processing using industrial scalable technologies such as cross-flow filtration and centrifugation for concentration prior to freezing or lyophilization of the cells and shelf-life stability testing.

We have successfully up-scaled difficult to culture ‘Next generation’ probiotics (Akkermansia, F. Prausnitzi). This was done in our pilot plant where we offer:

• Upscaling aligned with final GMP requirements (contacts with CMO are established)
• 40L, 600L, 4500L bioreactor capacity in food-grade environment
• Flexible DSP options, including centrifugation and crossflow filtration integrated into the bioreactor allow for washing and concentrations of cells under fully anaerobic conditions
• Formulations (liquid, powder, encapsulated)
• (Accelerated) shelf-life testing
• Many types of packaging
• Quality checks for product release of products

Tech-Transfer

We have experience with tech-transfer to CMO and GMP manufacturers of NIZO or jointly developed processes.