(228de) Crystallisation of Intact Monoclonal Antibody anticd20

Silica based nanotemplates with tuneable mesoporosity and surface chemistry, have been synthesised to facilitate protein crystallisation.¹ These nucleants act as a substrate for nucleation for macromolecular monoclonal antibodies (mAbs), which have greater complexity and segmental flexibility than many proteins. The class of anti cd20 mAbs is one of the most important therapeutics used in the treatment of lymphoproliferative disorders in the last 30 years.² Rituximab is a well known and widely used anti cd20. It has been crystallised previously both intact and as mAb fragments. Unfortunately, regardless of how similar anti cd20 mAbs may be, not all crystallise under the same solutions conditions.

Porous nanotemplates, tuned to the appropriate size, have been used to crystallise mAb anti cd20 by creating a spherulite seed stock. Templates have shown to significantly increase the number of crystals produced compared to crystallisation void of the heterogeneous nucleants. Stock solutions of these seeds were used for crystallisation and showed significant improvement in crystal quality. Moreover, seeding in different crystallisation conditions lead to various crystals with different morphologies.

References:

1. U.V. Shah, D.R. Williams, and J.Y.Y. Heng, â??Selective Crystallization of Proteins Using Engineered Nanonucleantsâ?,Cryst. Growth Des.,(2012), 12(3), 1362â??1369.
2. Lim, S.H., et al., Anti-CD20 monoclonal antibodies: historical and future perspectives. haematologica, 2010. 95(1): p. 135-143.