One of the crucial steps in deploying plasma therapy for COVID-19 is the development of critical assays, including ELISA, to detect SARS-CoV-2 antibodies and evaluate their potential suitability. Unfortunately, some assays for assessing neutralizing antibodies involve using live SARS-CoV-2 virus in a high containment facility (BSL3) and can take up to a week to complete. The assay cost, time for completion, and requirement for PPE make it a weak link in the pipeline for identifying plasma donors. Current commercial ELISAs provide primarily a yes/no answer and lack the precise quantitation of protective antibody levels desired in sourcing plasma. Therefore, the goal of this project is to biomanufacture and rapidly deploy coronavirus antigens in order to scale up a quantitative ELISA assay for characterizing plasma needed for treatment and prophylaxis. Our team is developing methodologies to generate purified SARS-CoV spike (S) protein and the SARS-CoV-2 S protein receptor binding domain (S-RBD) to be incorporated into the quantitative ELISA assay. The presence of an extremely sensitive and robust ELISA assay using multiple antigens will specify the highest binding and protective antibodies (and their targets) from a pool of volunteers and recipients. Additionally, the team is collaborating to develop an automated ELISA robotic system to increase efficiency and speed, ensuring reproducibility when performing candidate screening. This presentation will discuss the production of multiple S and RBD protein reagents using various production and purification platforms and the evaluation of their effectiveness through quantitative ELISA kits.

Biomanufacturing and implementation of sensitive quantitative COVID-19 ELISA platform to identify Sera needed for plasma therapy

ACS Fall 2021

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