RII Track-2 FEC: Membrane Purification Platform for Continuous Biomanufacturing of Viral Vectors and Virus-like Particles in Arkansas and Beyond

生物制剂是一种大分子疗法，已被用于治疗各种疾病，包括癌症、糖尿病、心血管疾病、自身免疫性疾病、遗传和眼科疾病等。生物制剂包括单抗(MAbs)和其他治疗性蛋白质、基因疫苗等疫苗、干细胞和病毒载体等细胞和基因治疗产品。生物制品对人类健康的影响是巨大的，新冠肺炎基因疫苗的成功就是一个例子。生物制品市场正在呈指数级增长，截至目前已达到3000亿美元以上。对生物制品的巨大需求对生产过程提出了巨大的挑战。下游工艺的总成本高达总制造成本的80%。RII Track-2 FEC项目的愿景是为未来大规模的病毒载体和病毒样颗粒(VLP)的生物制造创造一个基于膜的下游纯化平台，用于基因治疗和疫苗应用。该平台将被用来取代目前难以扩大规模的离心和树脂层析等过程。该项目的总体目标是开发创新和实用的单元操作，这些操作可以成为这些病毒颗粒疗法下游纯化平台的一部分。该项目汇集了来自阿肯色大学(阿肯色州)、克莱姆森大学(南卡罗来纳州)和肯塔基州大学(肯塔基州)三个EPSCoR辖区的具有互补专业知识的多学科研究人员。这三个机构中的每一个都提供了独特的设施和资源，可以迅速促进技术发展。拟议的活动对整个阿肯色州以及肯塔基州和南卡罗来纳州的经济影响将是巨大的。未来生物技术产业区域孵化中心将在EPSCoR的三个州建立。高生产率膜净化平台的成功开发预计将改善中低收入家庭获得革命性遗传和慢性病治疗的机会，从而改善人类健康。未来生物技术行业的劳动力发展，通过与少数族裔服务机构的合作致力于多样化，以及专注于职业生涯早期的教师指导是该项目的关键组成部分。该项目将为研究生提供实习机会，为本科生提供研究经验，以发展这一领域的STEM劳动力。此外，所有三所大学都将与该州各自的少数民族服务机构进行接触，以培养合作研究经验。发展经济高效的大规模生物制造来纯化病毒载体和病毒样颗粒(VLP)是一个主要的挑战。基于膜的单元操作很有吸引力；然而，膜容量、污染和缺乏商用膜，以及在一系列操作条件下膜性能的稳定性限制了工业从业者的应用。该项目将解决关键问题并克服关键障碍，为病毒载体和VLP开发基于膜的生物处理平台。该项目旨在克服多方面的技术和经济障碍，并涉及三个EPSCoR管辖区三个不同机构的研究人员之间的密切合作。更具体的研究目标包括(1)用于基因治疗的两种常见病毒载体和用于疫苗应用的VLP的原料生产；(2)用于生物反应器收获的先进微滤操作；(3)高容量亲和膜的设计、制造和表征；(4)用于分离全病毒衣壳和空病毒衣壳的下游膜层析；(5)组件设计、流程优化和流程强化；(6)使用最先进的生物分析方法进行检测和定量；以及(7)技术准备和验收研究。通过该项目在三所大学的合作研究，研究人员将在每个机构现有基础设施的基础上进行建设，远远超出目前的能力。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Biologics are large molecule therapeutics and have been used to treat a variety of diseases, including cancer, diabetes, cardiovascular disease, autoimmune disorders, genetic and ophthalmologic diseases to name a few. Biologics include monoclonal antibodies (mAbs) and other therapeutic proteins, vaccines such as mRNA vaccines, cell and gene therapies products such as stem cells, and viral vectors. The impact of biologics on human health has been tremendous with the success of mRNA vaccines for COVID-19 as an example. The market for biologics is increasing exponentially, reaching over $300 billion as of today. The large demand for biologics places great challenge on the manufacturing process. The total cost for downstream process accounts for up to 80% of the total manufacturing cost. The vision of this RII Track-2 FEC project is to create a membrane-based downstream purification platform for the future large scale biomanufacturing of viral vectors and virus-like particles (VLPs) for gene therapy and vaccine applications. This platform will be used to replace the current processes such as centrifugation and resin-based chromatography that are difficult to scale up. The overall goal of this project is to develop innovative and practical unit operations that could be part of a platform for the downstream purification of these virus particle-based therapeutics. The project brings together multidisciplinary researchers with complementary expertise from three EPSCoR jurisdictions from the University of Arkansas (Arkansas), Clemson University (South Carolina) and University of Kentucky (Kentucky). Each of the three institutions offers unique facilities and resources that can rapidly catalyze the technology development. The economic impact of the proposed activities on the entire state of Arkansas as well as on Kentucky and South Carolina will be significant. Regional incubator centers for biotechnology industry of tomorrow will be established in the three EPSCoR states. Successful development of a high-productivity membrane purification platform is expected to improve human health by improving access to revolutionary treatments for genetic and chronic diseases for mid- and low-income families. Workforce development for the future biotechnology industry, a commitment to diversity through collaborations with Minority-serving Institutions, and a focus on early career faculty mentorship are key components of the project. The project will provide internship opportunities for graduate students and research experiences for undergraduates to develop the STEM workforce in this field. In addition, all three universities will outreach to the respective Minority-serving Institutions in the state to foster collaborative research experiences. Development of cost-effective large scale biomanufacturing for the purification of viral vectors and virus-like particles (VLPs) is a major challenge. Membrane based unit operations are attractive; however, membrane capacity, fouling and lack of commercial membranes as well as the robustness of membrane performance under a range of operating conditions have limited applications by industrial practitioners. This project will address key issues and overcome critical barriers for developing a membrane-based bioprocessing platform for viral vectors and VLPs. The project aims to overcome the technical and economic barriers at multiple fronts and involve close collaborations among researchers at three different institutions in three EPSCoR jurisdictions. More specific research objectives include (1) feedstock production of two common viral vectors for gene therapy applications and VLPs for vaccine applications; (2) advanced microfiltration operations for bioreactor harvesting; (3) high capacity affinity membrane design, fabrication and characterization; (4) downstream membrane chromatography for separating full and empty viral capsids; (5) module design, process optimization and process intensification; (6) employing state-of-the-art bioanalytical methods for detection and quantification; and (7) a technology readiness and acceptance study. Through the project's collaborative research efforts across three universities, researchers will build upon existing infrastructure at each institution, expanding it far beyond current capabilities.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.