A scalable, integrated downstream process for production of a recombinant measles virus-vectored vaccine. Issue 9 (23rd February 2022)

• Record Type: Journal Article
• Authors: Steppert, Petra; Mosor, Magdalena; Stanek, Larissa; Burgstaller, Daniel; Palmberger, Dieter; Preinsperger, Shirin; Pereira Aguilar, Patricia; Müllner, Matthias; Csar, Patrick; Jungbauer, Alois
• Is Part Of: Vaccine. Volume 40:Issue 9(2022)
• Page Start: 1323
• Page End: 1333
• Publication Date: 2022-02-23
• Subjects: Infectious virus purification -- Tangential flow filtration -- Flow-through chromatography -- Membrane fouling -- Direct loading; Vaccines -- Periodicals; 615.372
• DOI: 10.1016/j.vaccine.2022.01.004
• Languages: English
• ISSNs: 0264-410X

Abstract

Highlights: A robust purification process for larger viruses such the Paramyxoviridae family, like measles. The order of process steps influences measles virus purity and titer. Process consists of restricted access chromatography and ultrafiltration. Process is suited for operation in a functional closed system under aseptic conditions. Scaleability to pilot scale level is proven. Abstract: Purification of very large and complex, enveloped viruses, such as measles virus is very challenging, it must be performed in a closed system because the final product cannot be sterile filtered and often loss of virus titer and poor product purity has been observed. We developed a purification process where the clarified and endonuclease treated culture supernatant is loaded on a restricted access chromatography medium where small impurities are bound and the virus is collected in the flow-through, which is then concentrated, and buffer exchanged by ultra/diafiltration. Up to 98.5% of host cell proteins could be captured by direct loading of clarified and endonuclease treated cell culture supernatant. Reproducible process performance and scalability of the chromatography step were demonstrated from small to pilot scale, including loading volumes from 50 mL up to 9 L. A 10-fold virus concentration was achieved by the ultrafiltration using a 100 kDa flat-sheet membrane. The order of individual process steps had a large impact on the virus infectivity and total process yields. The developed process maintained virus infectivity and is twice as fast as the traditional process train, where concentration is performed before loading on the chromatography column. Capturing impurities by the restricted access medium makes it a platform purification process with a high flexibility, which can be easily and quickly adapted to other vectors based on the measles virus vector platform.