Crystallization of a nonreplicating rotavirus vaccine candidate. Issue 4 (19th February 2021)

• Record Type: Journal Article
• Authors: Hong, Moo Sun; Kaur, Kawaljit; Sawant, Nishant; Joshi, Sangeeta B.; Volkin, David B.; Braatz, Richard D.
• Is Part Of: Biotechnology and bioengineering. Volume 118:Issue 4(2021)
• Page Start: 1750
• Page End: 1756
• Publication Date: 2021-02-19
• Subjects: crystallization modeling -- protein crystallization -- rotavirus -- vaccine development; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.27699
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Nonreplicating rotavirus vaccine (NRRV) candidates are being developed with the aim of serving the needs of developing countries. A significant proportion of the cost of manufacturing such vaccines is the purification in multiple chromatography steps. Crystallization has the potential to reduce purification costs and provide new product storage modality, improved operational flexibility, and reduced facility footprints. This communication describes a systematic approach for the design of the crystallization of an NRRV candidate, VP8 subunit proteins fused to the P2 epitope of tetanus toxin, using first‐principles models and preliminary experimental data. The first‐principles models are applied to literature data to obtain feasible crystallization conditions and lower bounds for nucleation and growth rates. Crystallization is then performed in a hanging‐drop vapor diffusion system, resulting in the nucleation and growth of NRRV crystals. The crystals obtained in a scaled‐up evaporative crystallization contain proteins truncated in the P2 region, but have no significant differences with the original samples in terms of antibody binding and overall conformational stability. These results demonstrate the promise of evaporative crystallization of the NRRV. Abstract : The crystallization of an nonreplicating rotavirus vaccine (NRRV) candidate was designed by a systematic approach using first‐principles models and preliminary experimental data. The first‐principles models are applied to literature data to obtain feasible crystallization conditions and lower bounds for nucleation and growth rates. Proof‐of‐concept experiments resulted in the nucleation and growth of NRRV crystals which have no significant differences with the original samples in terms of antibody binding and overall conformational stability.