Jerry Heng Keynote
Seeding and Continuous Protein Crystallisation for Bioseparation
Huaiyu YANG, Wenqian CHEN and Jerry Y. Y. HENG
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It is well known that nucleation is a conception stage and governs the entire crystallisation process. In this work, the influence of porosity interfaces on heterogeneous nucleation and crystallisation of macromolecules, the influence of shear rate distributions on nucleation behaviors in continuous crystallisation crystallisers has been investigated.
A Template Induced Polymorphic Occurrence Domain (TiPOD) concept on the nucleation of different polymorphic forms for small organic molecules is presented with the experimental results, demonstrating the role of porosity on the nucleation and crystallisation of macromolecules. The influence of seeding on the induction times for a batch crystallisation process and influence of seeding for bioseparation have been tested. An approach to detect surface heterogeneous nucleation based on Quartz Crystal Microbalance (QCM) will also be discussed. Computational fluid dynamics (CFD) simulations show increases in the amplitudes and frequencies accelerate the velocity of the oscillatory flows of the solution in the crystalliser, and correspondingly enhance the shear rate. The induction time of same system in oscillatory flow crystalliser (OFC) with equal driving force increases with increasing amplitude and frequency, then increases with a further increase of the frequency and amplitude, due to the complex influence of shear rate on kinetics of nucleation.
The work here demonstrates the importance of surface chemistry and surface porosity on heterogeneous nucleation and crystallisation and the importance of shear rate on the nucleation. Both template crystallisation and continuous crystallisation are developing fast and a potential more efficient and automatic method for manufacturing and purifying. By knowing the influence of porosity surface and kinetics on nucleation, we can control the crystallisation process. Further, we demonstrate the potential of a continuous protein crystallisation platform for downstream separation.
Acknowledgements: This work is funded by an EPSRC Manufacturing the Future Fellowship (EP/N015916/1) (JH) and AMECRYS is funded by the EU Horizon 2020 research and innovation programme under grant agreement no. 712965. Reference: [1] J. V. Parambil, S. K. Poornachary, S. J. Hinder, R. B. H. Tan, J. Y. Y. Heng, “Establishing template-induced polymorphic domains for API crystallisation: The case of Carbamazepine”, CrystEngComm (2015), 17, 6384-6392. [2] Yang, H.; Song, C. L.; Lim, Y. X. S.; Chen, W.; Heng, J.; (2017) “Selective crystallisation of carbamazepine polymorphs on differing surface properties”, CrystEngComm, DOI - 10.1039/C7CE01317E [3] U. V. Shah, D. R. Williams and J. Y. Y. Heng, “Selective crystallisation of proteins using engineered nanonucleants”, Cryst. Growth Des. (2012), 12, 1362-1369. [4] J.V. Parambil, M. Schaepertoens, D.R. Williams and J.Y.Y. Heng, “Effects of intermittent flow on the nucleation and crystallisation of insulin”, Cryst. Growth Des. (2011), 11, 4353-4359. [5] U. V. Shah, C. Amberg, Y. Diao, Z. Yang, J. Y. Y. Heng, “Heterogeneous nucleants for crystallogenesis and bioseparation”, Curr. Opin. Chem. Eng. (2015), 8, 69-75. [6] D. Wang, Z. Da, B. Zhang, M.A. Isbell, Y. Dong, X. Zhou, H. Liu, Z. Yang, J. Y. Y. Heng, “The stability study of tubular DNA origami in the presence of protein crystallisation buffer”, RSC Adv. (2015), 5, 58734-58737. [7] W. N. Al Nasser, U. V. Shah, K. Nikiforou, P. Petrou, J. Y. Y. Heng, “Effect of silica nanoparticles to prevent calcium carbonate scaling using an in situ turbidimetre”, Chem. Eng. Res. Des. (2016), 110, 98-107. [8] T. Lapidot, K. L. S. Campbell, J. Y. Y. Heng, “Model for interpreting surface crystallization using quartz crystal microbalance: Theory and experiments”, Anal. Chem. (2016), 88, 4886-4893. [9] Yang, H.; Yu, X. ; Raval, V. ; Makkawi, Y.; Florence, A.; (2016) “The effect of oscillatory flow on nucleation kinetics of butyl paraben”, Cryst. Growth Des. 16, 875-886 |
