School of Chemical Engineering and Advanced Materials, The University of Adelaide, Adelaide, South Australia, Australia.
Global Life Sciences Solutions Australia Pty Ltd., Parramatta, New South Wales, Australia.
Biotechnol Bioeng. 2022 Aug;119(8):2122-2133. doi: 10.1002/bit.28118. Epub 2022 May 10.
In this study, we present the first integrated and continuous downstream process for the production of microbial virus-like particle vaccines. Modular murine polyomavirus major capsid VP1 with integrated J8 antigen was used as a model virus-like particle vaccine. The integrated continuous downstream process starts with crude cell lysate and consists of a flow-through chromatography step followed by periodic counter-current chromatography (PCC) (bind-elute) using salt-tolerant mixed-mode resin and subsequent in-line assembly. The automated process showed a robust behavior over different inlet feed concentrations ranging from 1.0 to 3.2 mg ml with only minimal adjustments needed, and produced continuously high-quality virus-like particles, free of nucleic acids, with constant purity over extended periods of time. The average size remained constant between 44.8 ± 2.3 and 47.2 ± 2.9 nm comparable to literature. The process had an overall product recovery of 88.6% and a process productivity up to 2.56 mg h ml in the PCC step, depending on the inlet concentration. Integrating a flow through step with a subsequent PCC step allowed streamlined processing, showing a possible continuous pathway for a wide range of products of interest.
在这项研究中，我们提出了第一个用于生产微生物病毒样颗粒疫苗的集成式连续下游工艺。我们使用整合了 J8 抗原的模块化鼠多瘤病毒主要衣壳 VP1 作为模型病毒样颗粒疫苗。集成的连续下游工艺从粗细胞裂解物开始，包括流穿色谱步骤，随后使用耐盐混合模式树脂进行周期性逆流色谱（PCC）（结合-洗脱），以及随后的在线组装。该自动化工艺在不同的入口进料浓度（1.0 至 3.2 mg/ml）范围内表现出稳健的行为，仅需进行最小的调整，并且连续生产出高质量的病毒样颗粒，无核酸，纯度恒定，持续时间长。平均粒径在 44.8±2.3 和 47.2±2.9nm 之间保持不变，与文献相比无差异。该工艺的总产物回收率为 88.6%，在 PCC 步骤中的生产效率高达 2.56mg h ml，具体取决于入口浓度。将流穿步骤与随后的 PCC 步骤集成在一起，可以简化处理流程，为广泛的感兴趣产品展示了一种可能的连续途径。
