The use of virus-based biotherapeutics in the pharmaceutical industry have been progressed considerably in the last years, with applications in vaccination, gene therapy, and oncolytic therapy. However, given their biological complexity, there are still unmet challenges in the bioprocessing of these particles, particularly on the downstream processes. Insect-cell biopharmaceuticals produced using the baculovirus expression vector system (BEVS) have an additional purity-related concern due to the co-production of baculovirus particles. Their rod-shaped form and similar envelope to virus-like particles (VLPs) lead to a purification bottleneck, given the lack of efficiency in discriminating both particles.<br/>In the present work, a purification strategy based on an impurity-focused affinity chromatography approach for baculovirus removal from VLPs-based vaccine candidates manufacturing was developed. Several affinity ligands were discovered by phage display towards baculovirus particles and the most promising candidates were selected and immobilized in chromatographic support. The chromatographic performance and dynamic binding capacity were assessed for those resins. The impact of operational parameters, such as residence time was studied and their influence on the purification performance was described. The best candidates showed baculovirus removals above 70% and VLPs’ recoveries above 60%. As a proof-of-concept, these ligands were also evaluated using two different biopharmaceuticals produced by BEVS: hepatitis C VLPs and adeno-associated virus. Furthermore, these resins showed to be robust to the process and cleaning conditions, maintaining the purification performance over 20 cycles. Overall, by selecting the optimal operation setpoints, it was possible to develop a powerful chromatographic tool for baculovirus removal from biopharmaceuticals produced using BEVS.

Affinity-based approach for baculovirus removal from closely-related viral particles manufacturing

ACS Fall 2021

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