Inclusion Body Bead Size in E. coli Controlled by Physiological Feeding

摘要: The Gram-negative bacterium E. coli is the host of choice for producing a multitude recombinant proteins relevant in pharmaceutical industry. Generally, cultivation easy, media are cheap, and high product titer can be obtained. However, harsh induction procedures combined with usage IPTG (isopropyl β-d-1 thiogalactopyranoside) as an inducer often believed to cause stress reactions, leading intracellular protein aggregates, which so known so-called inclusion bodies (IBs). Downstream applications bacterial processes bottleneck overall process performance, bacteria lack many post-translational modifications, resulting time cost-intensive approaches. Especially purification notoriously its long processing times low yields. In this contribution, we present screening strategies determination body bead size coli-based bioprocess exclusively bodies. Size seen critical quality attribute (CQA), changes behavior have major effect on subsequent downstream processing. A model-based approach was used, aiming trigger distinct size: Physiological feeding control, using qs,C parameter, has impact could modelled hyperbolic saturation mechanism calculated form cumulated substrate uptake rate. Within model, sugar rate cells, uptake-value, simulated considered being key performance indicator desired size. We want highlight that mentioned strategy combination will allow tuning towards certain qs based control only. Optimized at time-point harvest should stabilize and, therefore, increase time-space yield. Furthermore, production may interesting application biocatalyst nanoparticulate material.