Historically, early stage E. coli clone screening has been carried out in shake flasks, but this method is time consuming and generally not predictive of bench scale cell line capabilities. To enhance the cell line selection process, a high throughput scale down model to identify the top performing clones for the expression of therapeutic proteins as inclusion bodies for large scale pharmaceutical production has been identified. The investigation has centered on the use of the Micro24 (Pall Corporation) to replace the shake flask model for the first round of clone screening. The design of an upstream process in the Micro24 has involved the optimization of closure methods, cassettes, batch volumes, feeding strategies and induction times to maximize protein expression and growth rate. Strategies to mitigate the high oxygen demand of E. coli and well to well process variations have also been characterized. In addition, the clone ranking from the Micro24 to the 1-L bench scale reactors has been compared. The study ultimately demonstrates that the Micro24, as a high-throughput platform, is a practical replacement for shake flasks with easy handling of 48-72 fermentations per week providing scalable clone selection for microbial projects in the biopharmaceutical setting.

Eliminating the shake flask: Developing the high throughput Micro24 platform for selecting an <i>E. coli</i> clone with scalable protein production

245th National Meeting (2013)

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Eliminating the shake flask: Developing the high throughput Micro24 platform for selecting an E. coli clone with scalable protein production