EAGER: SARS-CoV-2 Pseudovirus via a novel eukaryotic construction in yeast

抗击新冠肺炎这样的疾病的一个重大障碍是，无法获得一种安全有效的SARS-CoV-2病毒模仿物，这种病毒可以在不需要非常专门的操作和实验室设备的情况下使用，这些设备可以在技术开发过程中作为早期替代品。作为早期替代品，SARS-CoV-2模拟病毒或伪病毒可以作为安全、有效的工具来制造抗体、开发纯化方法、设计候选疫苗和建立诊断试剂盒。在缺乏合适的模仿的情况下，全国范围内快速通过/失败想法的能力非常有限。特别是，由于能够安全处理传染性病原体的设施数量很少，基础设施出现压力，最终限制了可能拥有丰富专业知识但住宿条件最低的团队的数量。该项目将产生这样一种安全的假病毒，可以广泛用于为这次和潜在的未来大流行开发材料。此外，博士后将接受跨学科环境的培训。这项建议的目标是开发一种合成酵母结构，作为SARS-CoV-2或其他传染病病原体的安全(BSL-1)、坚固和可靠的替代品，可以开发和验证诊断和环境测试分析。拟议工作的关键要素是1)开发酵母表面展示平台，在该平台中可以表达通常在SARS-CoV-2表面发现的关键蛋白质的表面密度，2)开发用于生物制造的强大的酵母伪病毒生产平台，以及3)针对SARS-CoV-2的酵母伪病毒的验证，以证明酵母伪病毒可以用作SARS-CoV-2的模拟物用于生物开发。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。

One of the significant impediments in the fight against a disease like COVID-19 is the availability of a safe and effective mimic of the SARS-CoV-2 virus that could be used without the need of very specialized handling and laboratory equipment that could serve as early-stage substitute during technology development. As an early stage substitute the SARS-CoV-2 mimic, or pseudovirus, could serve as a safe, effective tool to create antibodies, develop purification methods, design vaccine candidates, and build diagnostic kits. In the absence of suitable mimics, the nationwide ability to quickly pass/fail ideas is vastly limited. In particular, due to the low number of facilities that are able to safely handle an infectious agent, a strain on infrastructure occurs which ultimately limits the number of teams “in the fight” who may have significant expertise but minimal accommodations. This project will produce such a safe pseudovirus that could be widely used for the development of materials for this and potentially future pandemics. In addition, a postdoc will be trained in an interdisciplinary environment.The objective of this proposal is to develop a synthetic yeast construct that will serve as a safe (BSL-1), robust and reliable surrogate for SARS-CoV-2 or other infectious agents against which diagnostic and environmental testing assays could be developed and validated. The key elements of the proposed work are 1) the development of a yeast surface display platform in which the surface density of key proteins normally found on the surface of SARS-CoV-2 can be expressed, 2) the development of a robust production platform for the biomanufacturing of the yeast pseudovirus, and 3) the validation of the yeast pseudovirus against SARS-CoV-2 to demonstrate that the yeast pseudovirus can be used as a mimic for SARS-CoV-2 for the development of biologics.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.