Antibody Screening Results for Anti-Nucleocapsid Antibodies Toward the Development of a Lateral Flow Assay to Detect SARS-CoV-2 Nucleocapsid Protein

抗核衣壳抗体的抗体筛选结果,用于开发检测 SARS-CoV-2 核衣壳蛋白的横向流动检测法

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作者:David M Cate, Joshua D Bishop, Helen V Hsieh, Veronika A Glukhova, Luis F Alonzo, H Gleda Hermansky, Brianda Barrios-Lopez, Benjamin D Grant, Caitlin E Anderson, Ethan Spencer, Samantha Kuhn, Ryan Gallagher, Rafael Rivera, Crissa Bennett, Samantha A Byrnes, John T Connelly, Puneet K Dewan, David S B

Abstract

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.

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