There is a large unmet need for a prophylactic hepatitis C virus (HCV) vaccine to control the ongoing epidemic with this deadly pathogen. Many antiviral vaccines employ whole viruses as antigens. For HCV, this approach became feasible following the development of infectious cell culture systems for virus production. However, the lack of efficient downstream processes (DSP) for HCV purification poses a roadblock for the development of a whole virus vaccine. Using cell culture-derived genotype 1a HCV we developed a scalable and efficient DSP train, employing commonly used clarification and ultrafiltration techniques, followed by two membrane-based chromatography steps. For virus capture, steric exclusion chromatography using cellulose membranes was established, resulting in a virtually complete virus recovery with > 99% protein and 84% DNA depletion. Virus polishing was achieved by sulphated cellulose membrane adsorbers with ~ 50% virus recovery and > 99% protein and 90% DNA depletion. Additional nuclease digestion resulted in 99% overall DNA depletion with final DNA concentrations of 2 ng/mL. Process results were comparable for cell culture-derived HCV of another major genotype (5a). This study provides proof-of-concept for establishment of an efficient and economically attractive DSP with potential application for production of an inactivated whole virus vaccine against HCV for human use.

As a new initiative, HPV self-sampling to non-attenders using the dry Evalyn self-sampling brush is offered in the Capital Region of Denmark. The use of a dry brush is largely uncharted territory in terms of analytical stability. In this study we aim to provide evidence on the analytical quality of dry HPV self-sampling brushes as a function of time and temperature.We assessed the analytical stability of dry stored Evalyn brushes at three different temperatures, (4 °C, room temperature, 30 °C) and five different storage time points; T = 0 (baseline), 2, 4, 8, 16, and 32 weeks prior to HPV analysis using the BD Onclarity HPV assay.Mean Ct value of the Onclarity internal control was used as comparator of cellularity across time and temperatures, with no or only borderline statistical differences observed. HPV detection was stable throughout the five time points. In addition, analytically amplifiable DNA copy numbers and DNA fragmentation was assessed using the Agena iPLEX Exome QC assay, with no or only borderline statistical differences observed.In conclusion, the Evalyn brush is analytically stable with respect to human genomic material and HPV detection for up to 32 weeks at temperatures ranging from 4 °C to 30 °C.

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