TY - JOUR

T1 - Stability studies for the identification of critical process parameters for a pharmaceutical production of the Orf virus

AU - Eilts, Friederike

AU - Labisch, Jennifer J.

AU - Orbay, Sabri

AU - Harsy, Yasmina M.J.

AU - Steger, Marleen

AU - Pagallies, Felix

AU - Amann, Ralf

AU - Pflanz, Karl

AU - Wolff, Michael W.

N1 - Funding Information: The authors would like to thank Philip Wiese for the support with the infectivity analytics as well as Keven Lothert and Catherine Meckel-Oschmann for the proofreading of the manuscript. We kindly thank Albumedix Ltd. for the provided rHSA, Recombumin Prime, used in this study. The graphical abstract was created with biorender.com. This work was financially supported by the Heinrich-Böll-Foundation with a doctoral scholarship to F. Eilts. Additionally, an EXIST-Forschungstransfer grant (03EFKBW171) of the German Federal Ministry for Economic Affairs and Energy was granted and the project was co-funded by the European Regional Development Fund as part of the Union's response to the COVID-19 pandemic (IGJ-ERDF-Program Hesse - React EU 20008790). The presented manuscript is part of F. Eilts' dissertation at the Graduate Centre for Engineering Sciences under the aegis of the Justus Liebig University Giessen, Germany, in cooperation with the University of Applied Sciences Mittelhessen, Giessen, Germany.

PY - 2023/7/19

Y1 - 2023/7/19

N2 - A promising new vaccine platform is based on the Orf virus, a viral vector of the genus Parapoxvirus, which is currently being tested in phase I clinical trials. The application as a vaccine platform mandates a well-characterised, robust, and efficient production process. To identify critical process parameters in the production process affecting the virus’ infectivity, the Orf virus was subjected to forced degradation studies, including thermal, pH, chemical, and mechanical stress conditions. The tests indicated a robust virus infectivity within a pH range of 5–7.4 and in the presence of the tested buffering substances (TRIS, HEPES, PBS). The ionic strength up to 0.5 M had no influence on the Orf virus’ infectivity stability for NaCl and MgCl2, while NH4Cl destabilized significantly. Furthermore, short-term thermal stress of 2 d up to 37 °C and repeated freeze-thaw cycles (20 cycles) did not affect the virus’ infectivity. The addition of recombinant human serum albumin was found to reduce virus inactivation. Last, the Orf virus showed a low shear sensitivity induced by peristaltic pumps and mixing, but was sensitive to ultrasonication. The isoelectric point of the applied Orf virus genotype D1707-V was determined at pH 3.5. The broad picture of the Orf virus’ infectivity stability against environmental parameters is an important contribution for the identification of critical process parameters for the production process, and supports the development of a stable pharmaceutical formulation. The work is specifically relevant for enveloped (large DNA) viruses, like the Orf virus and like most vectored vaccine approaches.

AB - A promising new vaccine platform is based on the Orf virus, a viral vector of the genus Parapoxvirus, which is currently being tested in phase I clinical trials. The application as a vaccine platform mandates a well-characterised, robust, and efficient production process. To identify critical process parameters in the production process affecting the virus’ infectivity, the Orf virus was subjected to forced degradation studies, including thermal, pH, chemical, and mechanical stress conditions. The tests indicated a robust virus infectivity within a pH range of 5–7.4 and in the presence of the tested buffering substances (TRIS, HEPES, PBS). The ionic strength up to 0.5 M had no influence on the Orf virus’ infectivity stability for NaCl and MgCl2, while NH4Cl destabilized significantly. Furthermore, short-term thermal stress of 2 d up to 37 °C and repeated freeze-thaw cycles (20 cycles) did not affect the virus’ infectivity. The addition of recombinant human serum albumin was found to reduce virus inactivation. Last, the Orf virus showed a low shear sensitivity induced by peristaltic pumps and mixing, but was sensitive to ultrasonication. The isoelectric point of the applied Orf virus genotype D1707-V was determined at pH 3.5. The broad picture of the Orf virus’ infectivity stability against environmental parameters is an important contribution for the identification of critical process parameters for the production process, and supports the development of a stable pharmaceutical formulation. The work is specifically relevant for enveloped (large DNA) viruses, like the Orf virus and like most vectored vaccine approaches.

KW - Downstream processing

KW - Forced degradation studies

KW - Formulation

KW - Parapox Orf virus

KW - Viral vector vaccine

UR - http://www.scopus.com/inward/record.url?scp=85162863385&partnerID=8YFLogxK

U2 - 10.1016/j.vaccine.2023.06.047

DO - 10.1016/j.vaccine.2023.06.047

M3 - Article

C2 - 37353451

AN - SCOPUS:85162863385

VL - 41

SP - 4731

EP - 4742

JO - Vaccine

JF - Vaccine

SN - 0264-410X

IS - 32

ER -