DEVELOPMENT OF A NOVEL HIGH-THROUGHPUT PLATFORM FOR EFFICIENT PERFUSION-BASED CELL CULTURE PROCESS DEVELOPMENT

摘要

Perfusion technology has been successfully used for the commercial production of biotherapeutics, in particular unstable recombinant proteins, for more than a decade. However, there has been a general lack of high-throughput cell culture tools specifically for perfusion-based cell culture processes. Here we have developed a high-throughput cell retention operation for use with the ambr® 15 bioreactor system. Metabolite, titer, and cell concentrations were modeled using Matlab software and process parameters were defined to mimic continuous perfusion systems, with experimental results confirming model fidelity. Employing an offline process for cell retention and a variable volume approach, the established platform has demonstrated cell culture performance (> 90 E6 cells/mL and >95% viability) comparable to bench scale bioreactors. The model was successfully implemented for use in media development and clone screening with similar ranking of clones across scales, highlighting the accuracy of a microreactor screening approach and the utility of the ambr® 15 perfusion mimic. The automated, single use, high-throughput perfusion platform is a powerful tool that allows for rapid and efficient process development and demonstrates significant improvements over other established methods.