洞察到产品和流程相关的挑战的lentiviral 载体生物处理
Christopher Perry1,2,3, Noor Mujahid1, Yasu Takeuchi2,3
1Department of Biochemical Engineering, University College London, London, UK.
Biotechnology and bioengineering
|August 1, 2023
概括
这项研究证明了跨制造步骤的lentiviral vector (LV) 可加工性,揭示了特定的伪型在高剪切和盐度下是坚固的,与常见假设相反.
科学领域:
- 生物技术是生物技术.
- 基因治疗 基因治疗
- 过程开发 过程开发
背景情况:
- 长病毒载体 (LVs) 对于基因疗法至关重要,使长期基因表达成为可能.
- 优化LV生产需要了解他们在生物处理过程中的行为.
- 不同的LV伪型对制造工艺的影响尚未得到充分证实.
研究的目的:
- 在下游制造过程中评估不同晶状病毒载体伪型的可加工性.
- 研究制造条件 (温度,剪切,膜化学) 对LV回收的影响.
- 找出改善LV制造业收益率的机会.
主要方法:
- 在不同的制造条件下对三个常见的LV包膜蛋白进行比较.
- 在澄清和触流过 (TFF) 过程中评估载体恢复.
- 超缩放技术用于模拟高剪切环境并优化阳离子交换染色学.
主要成果:
- 过器膜化学显著影响载体恢复,取决于伪型.
- 在高剪切制造环境中,LVs表现出完全的稳定性.
- 在TFF模仿过过程中较高的剪切率增加了载体恢复.
- 开发了单体格式的优化离子交换染色体.
- LVs对高度盐 (高达1.7M) 的耐受性显示.
结论:
- 病毒载体伪型在下游制造过程中显著影响加工能力和恢复.
- 与普遍认为的相反,在高剪切和盐的条件下,LV是坚固的.
- 优化制造参数,如剪切率和色谱,可以提高LV回收和生产效率.
相关概念视频
01:27Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
01:23Vaccine Production
Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...