In this work, we investigated a unit operation from the past (flocculation) with the aim to make sense of the complexity of its interaction with centrifugation using a new approach (ultra scale-down studies) and in combination with novel data analysis techniques. The use of flocculation in primary recovery enhances the efficiency of clarification and product isolation by centrifugation. Flocculation is an important recovery operation for many biological products and yet its process development is often challenging. It is a complex unit operation which can be affected by operational conditions as well as bioprocess materials properties. To facilitate rapid bioprocess development, an ultra-scale down (USD) flocculation system was created to be used in tandem with the kompAsTM USD centrifugation. This allows studies to be undertaken at high throughput using millilitre quantities of material (as little as 50 mL). Consequently, relatively large amounts of data can be easily generated. Thus, multivariate data analysis techniques such as Principal Component Analysis and novel floc strength index (FSI’s) technique were integrated as a platform to handle the complex data and assess floc characteristics. The aim of this oral presentation is to show step-by-step how to develop a USD platform to enhance and select primary recovery options. This study used homogenate of E.coli expressing a relevant industrial enzyme to show that USD systems can mimic pilot scale performance and product yield. The novel FSI’s were used to easily characterise the flocs and inform centrifugation performance. At the end, this platform was validated at pilot scale using different flocculation conditions. The results show that pilot scale and USD platform produced results which are not statistically different. This suggests that these USD tools can be used to inform primary recovery performance. The ultra scale-down techniques developed allowed us to rapidly study several process conditions and enabled us to gain process understanding (mainly to focus on flocculation conditions that will improve protein yield while maintaining centrifugation performance). The use of MVDA for data evaluation provided easier data interpretation and visualisation of the relationships within a complex set of data. The ultra scale-down platform linked with advanced data analysis establishes a framework for future bioprocess studies to gain insights for enhanced primary recovery.

Step-by-step: How to develop a scale-down platform of flocculation and centrifugation for enhanced primary recovery

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.