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Open AccessArticle

Optimization of Vapor Diffusion Conditions for Anti-CD20 Crystallization and Scale-Up to Meso Batch

1
Department of Chemical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
2
Istituto di Cristallografia, CNR, via Amendola 122/o, Bari 70126, Italy
3
Istituto per la Tecnologia delle Membrane, CNR, Via P. Bucci Cubo 17C, Rende 87036, Italy
*
Authors to whom correspondence should be addressed.
Crystals 2019, 9(5), 230; https://doi.org/10.3390/cryst9050230
Received: 12 March 2019 / Revised: 18 April 2019 / Accepted: 24 April 2019 / Published: 29 April 2019
(This article belongs to the Special Issue Protein Crystallography)
The crystal form is one of the preferred formulations for biotherapeutics, especially thanks to its ability to ensure high stability of the active ingredient. In addition, crystallization allows the recovery of a very pure drug, thus facilitating the manufacturing process. However, in many cases, crystallization is not trivial, and other formulations, such as the concentrate solution, represent the only choice. This is the case of anti-cluster of differentiation 20 (anti-CD20), which is one of the most sold antibodies for therapeutic uses. Here, we propose a set of optimized crystallization conditions for producing anti-CD20 needle-shaped crystals within 24 h in a very reproducible manner with high yield. High crystallization yield was obtained with high reproducibility using both hanging drop vapor diffusion and meso batch, which is a major step forward toward further scaling up the crystallization of anti-CD20. The influence of anti-CD20 storage conditions and the effect of different ions on the crystallization processes were also assessed. The crystal quality and the high yield allowed the first crystallographic investigation on anti-CD20, which positively confirmed the presence of the antibody in the crystals. View Full-Text
Keywords: anti-CD20; crystallization; X-ray diffraction anti-CD20; crystallization; X-ray diffraction
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Yang, H.; Belviso, B.D.; Li, X.; Chen, W.; Mastropietro, T.F.; Di Profio, G.; Caliandro, R.; Heng, J.Y.Y. Optimization of Vapor Diffusion Conditions for Anti-CD20 Crystallization and Scale-Up to Meso Batch. Crystals 2019, 9, 230.

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