Cate David M, Bishop Joshua D, Hsieh Helen V, Glukhova Veronika A, Alonzo Luis F, Hermansky H Gleda, Barrios-Lopez Brianda, Grant Benjamin D, Anderson Caitlin E, Spencer Ethan, Kuhn Samantha, Gallagher Ryan, Rivera Rafael, Bennett Crissa, Byrnes Samantha A, Connelly John T, Dewan Puneet K, Boyle David S, Weigl Bernhard H, Nichols Kevin P
Global Health Labs, 14360 SE Eastgate Way, Bellevue, Washington 98007, United States.
Intellectual Ventures Laboratory, 14360 SE Eastgate Way, Bellevue, Washington 98007, United States.
ACS Omega. 2021 Sep 21;6(39):25116-25123. doi: 10.1021/acsomega.1c01253. eCollection 2021 Oct 5.
The global COVID-19 pandemic has created an urgent demand for large numbers of inexpensive, accurate, rapid, point-of-care diagnostic tests. Analyte-based assays are suitably rapid and inexpensive and can be rapidly mass-produced, but for sufficiently accurate performance, they require highly optimized antibodies and assay conditions. We used an automated liquid handling system, customized to handle arrays of lateral flow (immuno)assays (LFAs) in a high-throughput screen, to identify anti-nucleocapsid antibodies that will perform optimally in an LFA. We tested 1021 anti-nucleocapsid antibody pairs as LFA capture and detection reagents with the goal of highlighting pairs that have the greatest affinity for the nucleocapsid protein of SARS-CoV-2 within the LFA format. In contrast to traditional antibody screening methods (e.g., ELISA, bio-layer interferometry), the method described here integrates real-time reaction kinetics with transport in, and immobilization directly onto, nitrocellulose. We have identified several candidate antibody pairs that are suitable for further development of an LFA for SARS-CoV-2.
全球新冠疫情引发了对大量廉价、准确、快速的即时诊断检测的迫切需求。基于分析物的检测方法速度足够快且成本低廉，能够迅速大规模生产，但为了实现足够准确的性能，它们需要高度优化的抗体和检测条件。我们使用了一种自动化液体处理系统，该系统经过定制，可在高通量筛选中处理横向流动（免疫）分析（LFA）阵列，以识别能在LFA中实现最佳性能的抗核衣壳抗体。我们测试了1021对抗核衣壳抗体对，将其作为LFA捕获和检测试剂，目的是找出在LFA形式下对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）核衣壳蛋白具有最大亲和力的抗体对。与传统抗体筛选方法（如酶联免疫吸附测定法、生物层干涉术）不同，本文所述方法将实时反应动力学与在硝酸纤维素上的传输以及直接固定相结合。我们已经鉴定出了几对抗体对，适合进一步开发用于SARS-CoV-2的LFA。
