The baffled microtiter plate: increased oxygen transfer and improved online monitoring in small scale fermentations.
Most experiments in screening and process development are performed in shaken bioreactors. Today, microtiter plates are the preferred vessels for small-scale microbial cultivations in high throughput, even though they have never been optimized for this purpose. To interpret the experimental results correctly and to obtain a base for a meaningful scale-up, sufficient oxygen supply to the culture liquid is crucial. For shaken bioreactors this problem can generally be addressed by the introduction of baffles. Therefore, the focus of this study is to investigate how baffling and the well geometry affect the maximum oxygen transfer capacity (OTR(max)) in microtiter plates. On a 48-well plate scale, 30 different cross-section geometries of a well were studied. It could be shown that the introduction of baffles into the common circular cylinder of a microtiter plate well doubles the maximum oxygen transfer capacity, resulting in values above 100 mmol/L/h (k(L)a > 600 1/h). To also guarantee a high volume for microbial cultivation, it is important to maximize the filling volume, applicable during orbital shaking. Additionally, the liquid height at the well bottom was examined, which is a decisive parameter for online-monitoring systems such as the BioLector. This technology performs fiber-optical measurements through the well bottom, therefore requires a constant liquid height at all shaking frequencies. Ultimately, a six-petal flower-shaped well geometry was shown to be the optimal solution taking into account all aforementioned criteria. With its favorable culture conditions and the possibility for unrestricted online monitoring, this novel microtiter plate is an efficient tool to gain meaningful results for interpreting and scaling-up experiments in clone screening and bioprocess development.
Funke
Matthias
M
AVT.Biochemical Engineering, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany.
Diederichs
Sylvia
S
Kensy
Frank
F
Müller
Carsten
C
Büchs
Jochen
J
D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods; D003470:Culture Media / Q000737:chemistry*; D005285:Fermentation; D010100:Oxygen / Q000378:metabolism*
D003470:Culture Media; D010100:Oxygen