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Microscale disruption of microorganisms for parallelized process development

09:38 EDT 21 Jun 2017 | Wiley Biotechnology Journal

Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris are the standard platforms for biopharmaceutical production with 40% of all between 2010 to 2014 approved protein drugs produced in those microbial hosts. Typically, products overexpressed E. coli and S. cerevisiae remain in the cytosol or are secreted into the periplasm. Consequently, efficient cell disruption is essential for high product recovery during microbial production. Process development platforms at microscale are essential to shorten time to market. While high‐pressure homogenization is the industry standard for cell disruption at large scale this method is not practicable for experiments in microscale. This review describes microscale methods for cell disruption at scales as low as 200 µL. Strategies for automation, parallelization and miniaturization, as well as comparability of the results at this scale to high pressure homogenization are considered as those criteria decide which methods are most suited for scale down. Those aspects are discussed in detail for protein overexpression in E. coli and yeast but also the relevance for alternative products and host such as microalgae are taken into account. The authors conclude that bead milling is the best comparable microscale method to large scale high‐pressure homogenization and therefore the most suitable technique for automated process development of microbial hosts with the exception of pDNA production. Cell disruption is crucial for protein recovery from many microbial hosts since those tend to dominantly overexpress recombinant products intracellular. For process development microscale methods are favorable which deliver homogenates which are highly comparable to those obtained with high pressure homogenization, the reference method for industrial scale cell disruption. Considering this aspect combined with available strategies for automation and parallelization microscale bead milling is the most suitable technique for automated process development of microbial hosts with the exception of pDNA production.

Original Article: Microscale disruption of microorganisms for parallelized process development

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