| Autor: |
Lerer E; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Oren Z; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Kafri Y; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Adar Y; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Toister E; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Cherry L; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Lupu E; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Monash A; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Levy R; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Dor E; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Epstein E; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Levin L; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Girshengorn M; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Natan N; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Zichel R; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel., Makovitzki A; Department of Biotechnology, Israel Institute for Biological Research (IIBR), Ness-Ziona 74100, Israel. |
| Abstrakt: |
This study reports a highly efficient, rapid one-step purification process for the production of the recombinant vesicular stomatitis virus-based vaccine, rVSV-∆G-spike (rVSV-S), recently developed by the Israel Institute for Biological Research (IIBR) for the prevention of COVID-19. Several purification strategies are evaluated using a variety of chromatography methods, including membrane adsorbers and packed-bed ion-exchange chromatography. Cell harvest is initially treated with endonuclease, clarified, and further concentrated by ultrafiltration before chromatography purification. The use of anion-exchange chromatography in all forms results in strong binding of the virus to the media, necessitating a high salt concentration for elution. The large virus and spike protein binds very strongly to the high surface area of the membrane adsorbents, resulting in poor virus recovery (<15%), while the use of packed-bed chromatography, where the surface area is smaller, achieves better recovery (up to 33%). Finally, a highly efficient chromatography purification process with Capto TM Core 700 resin, which does not require binding and the elution of the virus, is described. rVSV-S cannot enter the inner pores of the resin and is collected in the flow-through eluent. Purification of the rVSV-S virus with Capto TM Core 700 resulted in viral infectivity above 85% for this step, with the efficient removal of host cell proteins, consistent with regulatory requirements. Similar results were obtained without an initial ultrafiltration step. |
