(240e) Controversial Influence of Gravitational Force on the Crystallisation of Lysozyme

The manufacturing of biopharmaceuticals [1] in a cost effective and reliable route is a major industrial challenge due to the complex nature of the protein. Biocrystallisation [2,3] as an alternative to conventional downstream separations, offers route to efficient biopharmaceutical manufacturing, enabling higher throughput, lowering the product costs, and an increase in product quality and stability [4,5]. There are literatures reported the controversial results about the gravity, some reported micro-gravity enhanced the crystallisation of protein, and, however, some reported hyper-gravity accelerated the protein crystallisation.

In this work, the influence of gravity induced by centrifugation is investigated for variation in rotation speeds and time. This gravity effect is investigated for lysozyme crystallisation solution (70-90 mg/ml) with NaCl solution (90-110 mg/mL). The results obtained show no obvious influence of gravity force (< 100 g) and short duration (< 5 mins) of centrifugation on the crystallisation process. The results also answered the controversial results reported in literatures. For a continuously long time centrifugation, the high gravity (> 11000 g) hindered the nucleation process. However, the high gravity accelerated the crystal growth process, therefore, obtaining larger crystals comparing with the low gravity.The continuously manufacturing by combination of centrifugation and crystallisation can be an efficient platform for purifying macro-biomolecules.

Acknowledgements

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

References

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