Optimization of scalable chromatography-based purification methods utilizing NatriFlo HD-Q membrane technology for downstream processing of viral vaccines and viral vectors

病毒工程表达抗原，以促进对特定病原体的保护性免疫反应（疫苗），或提供治疗基因（基因疗法）或治疗癌症（溶瘤病毒疗法）是用于预防和治疗人类和动物疾病的基本工具。然而，使用病毒作为疫苗或治疗剂的一个不可克服的障碍是有效的、可扩展的纯化系统的可用性。该项目的目标是建立在我们最初与Natrix Separations进行NSERC Engage资助合作期间获得的工艺开发专业知识的基础上，在该合作中，我们成功开发了一种纯化高滴度临床级纽卡斯尔病毒的工艺，并将此知识应用于高滴度生产的详细工艺纯化方案的设计、优化和传播，使用NatriFlo HD-Q膜技术的临床级病毒疫苗和基因转移载体。NatrixSeparations是生命科学市场的高性能、一次性和多循环色谱产品的领先供应商。其膜色谱柱用于各种应用，包括肽、蛋白质、核酸和单克隆抗体的分离和纯化。Natrix正在寻求扩大其专利分离技术平台，以包括用于人类和动物的临床级病毒疫苗/载体和溶瘤病毒的纯化。虽然Natrix拥有色谱设备和工艺开发专业知识，但他们没有进行这些实验所需的基础设施（CL2实验室和动物设施，获得病毒）或病毒学专业知识。因此，Natrix与学术病毒学实验室合作开发病毒特异性纯化工艺并在动物模型中评估最终产品的活性和纯度绝对至关重要。事实上，这是我们最初的NSERC参与赠款的前提。由于这项合作的巨大成功，我们现在正在申请资金，以开发和测试用于各种临床相关病毒疫苗和病毒载体的阴离子交换和亲和色谱纯化工艺。

Viruses engineered to express antigens to promote a protective immune response against a particular pathogen(vaccine), or to deliver a therapeutic gene (gene therapy) or to treat cancer (oncolytic virotherapy) are essentialtools used in the prevention and treatment of diseases in humans and animals. However, a considerableimpediment to the use of viruses as vaccines or therapeutic agents is the availability of efficient, scalablepurification systems. The goal of this project is to build upon the process development expertise acquiredduring our initial NSERC Engage funded collaboration with Natrix Separations, in which we successfullydeveloped a process for purifying high titer, clinical grade Newcastle Disease virus, and apply this knowledgeto the design, optimization and dissemination of detailed process purification protocols for production of hightiter, clinical grade viral vaccines and gene transfer vectors using NatriFlo HD-Q membrane technology. NatrixSeparations is a leading supplier of high performance, single use and multi-cycle chromatography products tothe life science markets. Their membrane chromatography columns are used in a variety of applicationsincluding the separation and purification of peptides, proteins, nucleic acids, and monoclonal antibodies. Togrow their business, Natrix is seeking to broaden their patented separation technology platform to includepurification of clinical grade viral vaccines/vectors and oncolytic viruses for use in humans and animals. WhileNatrix has the chromatographic devices and process development expertise, they do not have the infrastructure(CL2 lab and animal facilities, access to viruses) or the virology expertise necessary to conduct theseexperiments. Therefore, it is absolutely critical that Natrix partner with an academic virology lab to developvirus-specific purification processes and evaluate the activity and purity of the final product in an animalmodel. Indeed, that was the premise of our initial NSERC Engage grant. Due to the tremendous success of thiscollaboration, we are now applying for funding to develop and test anion exchange and affinitychromatography-based purification processes for a variety of clinically relevant viral vaccines and viral vectors.