The purification of large viruses remains an important field of research and development. The development of efficient purification trains is restricted by limited analytical methods, as well as by the complexity of large viruses, as well as the high variability in starting material from cell culture. Vaccinia virus holds great potential as an oncolytic and immunotherapeutic vaccine against a broad spectrum of cancers. In this work, monolith-based capture and polishing chromatographic steps for vaccinia virus Lister strain has been developed. Virus produced in CV-1 cells was harvested and passed through a 0.8 μm pre-filter before loading onto CIEX, AIEX and HIC CIM monoliths. Without the need for nuclease treatment, up to 99% of the total DNA loaded can be removed from the vaccinia feed stream by the CIM OH monolith, which also reduces the total protein concentration in the product pool to LLOQ levels, and achieves infectious virus recoveries of 90%. Binding capacities of greater than 1 × 10⁹ pfu of vaccinia per mL of matrix were obtained on both CIM SO3 and CIM OH monoliths. Multiple orthogonal analytical methods have been used to develop process knowledge and understanding.

大病毒的纯化仍然是一个重要的研究和开发领域。有效纯化序列的开发受到有限的分析方法、大型病毒的复杂性以及细胞培养起始材料的高度可变性的限制。牛痘病毒作为一种针对广谱癌症的溶瘤和免疫治疗疫苗具有巨大的潜力。在这项工作中，针对牛痘病毒 Lister 株开发了基于整体的捕获和纯化色谱步骤。收集 CV-1 细胞中产生的病毒并通过 0.8 μm 预过滤器，然后加载到 CIEX、AIEX 和 HIC CIM 整体上。无需核酸酶处理，高达 99% 的总 DNA 负载可以通过 CIM OH 整体从牛痘进料流中去除，这还将产品池中的总蛋白质浓度降低到 LLOQ 水平，并实现 90% 的传染性病毒回收率。结合能力大于 1 × 10 在 CIM SO3 和 CIM OH 单块上均获得了每毫升基质^{9 pfu 的牛痘。}多种正交分析方法已被用于发展过程知识和理解。