(200i) Continuous Protein Crystallization of Lysozyme

The current work investigates the technical feasibility of performing continuous protein crystallisation in an oscillatory flow platform with lysozyme as the model protein. Different crystallisation conditions were generated by varying the protein concentration (30 to 100 mg/mL) as well as the oscillation amplitude (10 to 25 mm) and frequency (0.05 to 0.25 Hz). Based on the Reynolds number (Re) under different flow conditions, the shaking batch crystallisation was successfully transferred to the batch oscillatory flow crystallisation and subsequently the continuous oscillatory flow crystallisation. Experimental results show that the continuous crystallisation reached equilibrium state at a resident time of approximately 200 min and consistent crystal shape and size were achieved at the outlet of the crystalliser. This work presents a robust method for the development of continuous protein crystallisation in an oscillatory flow system, which is useful for the industrial downstream processing of proteins.^1,2

Acknowledgement

We acknowledge the EPSRC (EP/N015916/1) for funding.

Reference

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2. Yang, H., Yu, X., Raval, V., Makkawi, Y. & Florence, A. Effect of Oscillatory Flow on Nucleation Kinetics of Butyl Paraben. Cryst. Growth Des. 16, 875–886 (2016).