Single-use wave-mixed versus stirred bioreactors for insect-cell/BEVS-based protein expression at benchtop scale

Spodoptera frugiperda-9 (Sf-9) cells used in conjunction with the baculovirus expression vector system (BEVS) represent a promising platformfor the rapid development and manufacture of protein complexes and virus-like particle (VLP) products. Several studies have described the superiority of single-use wave-mixed bioreactors although reusable stirred and, more recently, single-use stirred bioreactors have also been successfully applied. Due to their bioengineering characteristics (more homogeneous energy dissipation, reduced foam formation), wave-mixed systems are often preferred. However, a direct comparison of the influence of single-use wave-mixed and single-use stirred bioreactors on cell growth and protein expression in Sf-9/BEVS-based production processes was still lacking. We investigated Sf-9 cell growth and expression of a recombinant secreted alkaline phosphatase (rSEAP) in thewave-mixed BIOSTAT® RMaswell as the stirred UniVessel R SU and a serum-free culture medium. Irrespective of the bioreactor system, comparable growth, substrate, and metabolite courses as well as peak cell densities (>1.2 × 107 cells mL−1) were observed in Sf-9 cell expansions performed in batch mode. Additionally, identical rSEAP quality and maximumrSEAP activities were found in biphasic productions in both bioreactor systems. Concluding, comparability of single-use wave-mixed and stirred bioreactors for insect cell culture processes was demonstrated for the first time.