Abstract
Recombinant protein and virus-like particle (VLP) production based on the baculovirus expression vector system is fast, flexible, and offers high yields. Independent from the product, a multitude of parameters are screened during process development/optimisation. Early development acceleration is a key requirement for economic efficiency, and µ-scale bioreactor systems represent an attractive solution for high-throughput (HTP) experimentation. However, limited practical knowledge is available on the relevance and transferability of screening data to pilot scales and manufacturing. The main goal of the present study was to evaluate a HTP µ-bioreactor platform with respect to its aptitude as a screening platform mainly based on transferability of results to benchtop bioreactors representing the conventional production regime. Second question was to investigate to what extent the online sensors of the µ-bioreactor contribute to process understanding and development. We demonstrated that transferability of infection screening results from the HTP µ-bioreactor scale to the benchtop bioreactor was equal or better than that from shaker cultivation. However, both experimental setups turned out to be sub-optimal solutions that only allowed for a first and rough ranking with low relevance in the case of absolute numbers. Bioreactor yields were up to one order of magnitude higher than the results of screening experiments.
Highlights
Recombinant protein and virus-like particle (VLP) production based on the baculovirus expression vector system is fast, flexible, and offers high yields
There are studies focused on development of mathematic models to calculate the best infection strategies including multiplicity of infection (MOI), TOI, concentration at time of infection (CCI) and media depletion[2,3,4]
The determination of virus titre is a critical and time-consuming step and, independent of the methods used, there is significant analytical error in the range of ± 1 log fold changes[9]. This is valid for both plaque assay and tissue culture infectious dose 50 (TCID50), two methods commonly used and accepted in academia and industry[10]
Summary
Recombinant protein and virus-like particle (VLP) production based on the baculovirus expression vector system is fast, flexible, and offers high yields. We demonstrated that transferability of infection screening results from the HTP μ-bioreactor scale to the benchtop bioreactor was equal or better than that from shaker cultivation Both experimental setups turned out to be sub-optimal solutions that only allowed for a first and rough ranking with low relevance in the case of absolute numbers. The determination of virus titre is a critical and time-consuming step and, independent of the methods used, there is significant analytical error in the range of ± 1 log fold changes[9] This is valid for both plaque assay and tissue culture infectious dose 50 (TCID50), two methods commonly used and accepted in academia and industry[10]. To identify the optimal CCI and MOI for high yield production of VLPs or proteins of interest, multiple costly and time-consuming cultivations have to be performed in small scale before transferring the process to a larger scale. ® flasks or cell culture flasks is the Biolector (m2p-labs GmbH, Baesweiler, Germany), a titre plate-based platform
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