We isolate B. fragilis from fecal and validated repositories under strict anaerobiosis (pO2 control, pre-reduced media, anaerobic transfer). Primary screens triage candidates on growth kinetics, acid/bile tolerance, and carbohydrate utilization. Parallel safety screens include bft gene PCR and hemolysis checks. Candidates advance on stability, genetic features, and preliminary metabolite profiles (e.g., acetate/propionate signatures).
Definitive classification integrates MALDI-TOF, 16S rRNA (full-length), and whole-genome sequencing to resolve clades and detect virulence/safety loci (e.g., bft, CPS clusters). We deliver annotated FASTA/GBK files, average nucleotide identity (ANI), and biosynthetic pathway calls relevant to capsular polysaccharides and polysaccharide utilization loci (PULs), enabling traceable, publication-ready IDs.
We engineer fed-batch or continuous anaerobic fermentations tuned for carbon flux (complex vs defined media), pH/ORP stability, and osmolality, optimizing biomass and metabolite yields. In-line analytics track OD, residual sugars, and off-gas. Downstream goals (viable cells vs cell-free fractions enriched in SCFAs) dictate agitation/feeding logic to minimize stress responses and maintain phenotype.
Cell harvest leverages centrifugation/filtration schemes compatible with obligate anaerobes. For metabolite fractions, we apply solvent-free clarification and validated filtration (MWCO selection) before targeted quantitation (GC-FID/MS for SCFAs). For cell products, oxygen-managed concentration and cryo-excipients preserve integrity ahead of stabilization and QC release.
We design research-grade formulations that combine dried B. fragilis powders with prebiotics, buffering systems, and moisture-protection strategies. These can be prepared into capsules, tablets, sachets, or liquid suspensions. Each format is optimized for rehydration performance, survival of viable cells, and usability, making them adaptable for different application pipelines.
Release testing spans identity (genomics/MALDI-TOF), purity (aerobe/anaerobe panels), endotoxin (LAL), CFU enumeration, and growth/fermentation kinetics. For safety research, we include bft PCR, BFT protein ELISA where applicable, and capsular polysaccharide profiling (PSA/PSB/PSC markers) to contextualize immune-active features.
We run in vitro readouts tied to B. fragilis biology: SCFA production rates, epithelial barrier modulation (TEER, junction proteins), and immune-crosstalk assays. Specialized tests quantify PSA-linked induction of IL-10-competent lymphocytes (co-culture with DC/T cells) vs ETBF-associated epithelial signaling (e.g., β-catenin pathway activation) to flag mechanistic directionality.
Human epithelial/immune co-cultures and organ-on-chip setups model mucosal exposure, cytokine patterns (IL-10, IL-8), and barrier dynamics. We implement TLR2-dependency checks for PSA studies and contrast ETBF vs non-toxigenic strains for pathway mapping (NF-κB, MAPK). Endpoint packages include multiplex cytokines, RNA-seq, and phospho-signaling panels.
