(469c) Continuous Crystallisation Oscillatory Flow Platform for Protein Purification and Bio-Separation
AIChE Annual Meeting
2020
2020 Virtual AIChE Annual Meeting
Separations Division
Continuous Crystallization Processes
Tuesday, November 17, 2020 - 8:30am to 8:45am
Here, we present a platform for continuous oscillatory flow crystallization (COFC) (Figure 1) and its application for the crystallization of lysozyme[1,2]. A workflow is developed from µl screening experiments, to scaled up batch oscillatory flow crystallization (BOFC) and COFC experiments.
The induction time for lysozyme is found to be inversely proportional to the initial protein concentration, with the same relationship found between the induction time and the frequency and amplitude of the oscillation. However, it is also observed that the decrease in induction time comes at a cost to the crystal size and quality. In turn, the high shear environment from increased amplitude and frequency of the COFC can also damage the fragile protein crystals, resulting in the formation of more fine particles and an undesirable size distribution. This led to further investigation of slug flow mixing in the continuous crystalliser, where the crystal size and shapes are compared to the particles produced from the osciallatory flow reactor. Further work around optimization of the crystallization parameters and crystallizer geometry using the gCrystal platform to find the optimum scale up conditions are also discussed. The use of seeding and potential templating materials is also covered with a view to expanding the use of this crystallizer to less well characterized protein materials.
It is demonstrated that purification of proteins can be achieved by crystallization in a continuous mode. This route offers the possibility to revolutionise downstream separations of biopharmaceuticals
References
[1] H.Y. Yang, W.Q. Chen, P. Peczulis, J.Y.Y. Heng (2019): Development and Workflow of a Continuous Protein Crystallization Process: A Case of Lysozyme, Cryst. Grow. Des; 19, 983-991.
[2] H.Y. Yang, P. Peczulis, P. Inguva, X.Y. Li, J.Y.Y. Heng (2018): Continous Protein Crystallisation Platform and Process: Case of Lysozyme, Chem. Eng. Res. Des. 136, 529-53