Development of an ELISA Assay for the Determination of SARS-CoV-2 Protein Subunit Vaccine Antigen Content

Autor: Lu Han, Chaoqiang An, Dong Liu, Zejun Wang, Lianlian Bian, Qian He, Jianyang Liu, Qian Wang, Mingchen Liu, Qunying Mao, Taijun Hang, Aiping Wang, Fan Gao, Dejiang Tan, Zhenglun Liang
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Viruses, Vol 15, Iss 1, p 62 (2022)
Druh dokumentu: article
ISSN: 1999-4915
DOI: 10.3390/v15010062
Popis: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) protein subunit vaccine is one of the mainstream technology platforms for the development of COVID-19 vaccines, and most R&D units use the receptor-binding domain (RBD) or spike (S) protein as the main target antigen. The complexity of vaccine design, sequence, and expression systems makes it urgent to establish common antigen assays to facilitate vaccine development. In this study, we report the development of a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to determine the antigen content of SARS-CoV-2 protein subunit vaccines based on the United States Pharmacopeia and ICH (international conference on harmonization) Q14 and Q2 (R2) requirements. A monoclonal antibody (mAb), 20D8, was identified as the detection antibody based on its high RBD binding activity (EC50 = 8.4 ng/mL), broad-spectrum anti-variant neutralizing activity (EC50: 2.7–9.8 ng/mL for pseudovirus and EC50: 9.6–127 ng/mL for authentic virus), good in vivo protection, and a recognized linear RBD epitope (369–379 aa). A porcine anti-RBD polyclonal antibody was selected as the coating antibody. Assay performance met the requirements of the analytical target profile with an accuracy and precision of ≥90% and adequate specificity. Within the specification range of 70–143%, the method capability index was >0.96; the misjudgment probability was
Databáze: Directory of Open Access Journals
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