Andaluz Angela, Monteverde Brittany, Vera Kevin, Tse Brandon, Gajic Ivan, Forelich Clifford, Motevalian Seyed Pouria
BioProcess Sciences, Pharma Services, Viral Vector Services, Thermo Fisher Scientific, Plainville, Massachusetts, USA.
Biotechnol Bioeng. 2025 Nov;122(11):3051-3060. doi: 10.1002/bit.70052. Epub 2025 Aug 21.
Adeno-associated virus (AAV) is one of the most common delivery systems used in gene therapy. Challenges in the development and manufacturing of AAVs include high cost of goods (COGs) per dose, process scalability, speed to market, and process-related impurities such as empty capsids. This article presents a streamlined approach to developing and scaling AAV upstream production process via triple transfection from bench scale to commercial volumes exceeding 1,000 L. By leveraging high-throughput technologies such as the AMBR®15 system, we achieved rapid upstream process development in under 2 months. These tools enabled optimization of productivity, impurity reduction, and COGs per dose. We also detail methodologies for direct scale-up from AMBR®15 to a 2,000 L single-use production bioreactor.
腺相关病毒（AAV）是基因治疗中最常用的递送系统之一。AAV开发和制造面临的挑战包括每剂商品成本（COG）高、工艺可扩展性、上市速度以及与工艺相关的杂质，如空衣壳。本文介绍了一种通过三次转染从实验室规模到超过1000升的商业规模简化AAV上游生产工艺开发和放大的方法。通过利用高通量技术，如AMBR®15系统，我们在不到2个月的时间内实现了快速的上游工艺开发。这些工具实现了生产力优化、杂质减少和每剂COG降低。我们还详细介绍了从AMBR®15直接放大到2000升一次性生产生物反应器的方法。
