Array Based Affinity Selection

项目摘要 需要资金来应用目前NIGMS奖正在开发的技术，以产生针对SARS-CoV-2病毒刺突蛋白的重组抗体样亲和试剂。作为一名 原理验证实验，我们已经分离出四个纤维连接蛋白III(FN3)单体 与病毒刺突蛋白的受体结合域(RBD)结合的纳摩尔亲和力低的蛋白。行政副刊将允许继续和扩大这项工作，以产生 与皮摩尔亲和力结合的亲和试剂，可用于敏感、可靠的诊断 唾液中病毒颗粒的分析(均相分析、侧向流动分析)。分开来看，这些 单体将被重新格式化为二价Fc融合，并在CHO细胞中过度表达。是这样的 试剂有可能作为治疗剂，降低或阻止病毒进入患者ACE2表达细胞。 1

Project Summary Funds are requested to apply technologies being developed in a current NIGMS award to generate recombinant antibody-like affinity reagents to the spike protein of the SARS-CoV-2 virus. As a proof-of-principle experiment, we have already isolated four fibronectin type III (FN3) monobodies that bind with low nanomolar affinity to the receptor binding domain (RBD) of the viral spike protein. An administrative supplement will permit continuation and expansion of this work to generate affinity reagents that bind with picomolar affinity and can be used in sensitive, robust diagnostic assays (homogenous assays, lateral flow assays) for virus particles in saliva. Separately, these monobodies will be reformatted as bivalent Fc fusions and overexpressed in CHO cells. Such reagents have the potential to serve as therapeutic agents that lower or block viral entry into patients ACE2-expressing cells. 1