Separating Empty and Full Recombinant Adeno-Associated Virus Particles Using Isocratic Anion Exchange Chromatography
| Autor: | John Pieracci, Christopher Argento, Meisam Bakhshayeshi, Ryan Dickerson |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Anions viruses Ion chromatography Genetic Vectors medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology law.invention law 010608 biotechnology medicine Adeno-associated virus Recombination Genetic Chromatography Ion exchange Elution Chemistry 010401 analytical chemistry Virion General Medicine Dependovirus Chromatography Ion Exchange 0104 chemical sciences Linear gradient Recombinant DNA Molecular Medicine Clinical safety Design space |
| Zdroj: | Biotechnology journal. 16(1) |
| ISSN: | 1860-7314 |
| Popis: | The development of recombinant adeno-associated virus (rAAV) gene therapies is becoming an increasing priority in the biotherapeutic landscape. One of the challenges associated with the production of rAAV is the formation of empty AAV particles that do not contain a therapeutic gene. The concerns about the impact of empty particles on clinical safety and rAAV-mediated gene expression have necessitated the development of purification processes to remove these species. The development of a robust and scalable purification process to separate empty and full AAV particles at large scale remains a challenge. In this study, a novel anion exchange chromatography process based on isocratic wash and elution steps to enrich full rAAV2 particles is presented. An operating design space is identified to ensure the robustness of the process. The isocratic chromatography provides several advantages over the traditional shallow linear gradient elution, including lower buffer consumption, smaller intermediate pool volumes, and more robust manufacturing. |
| Databáze: | OpenAIRE |
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