Chromatographic characterization of the fusion protein SARS-CoV-2 S protein (RBD)-hFc-Reference-Cited by-同舟云学术

Chromatographic characterization of the fusion protein SARS-CoV-2 S protein (RBD)-hFc

Published:2022-04-21 Issue:0 Volume:0 Page:
ISSN:2365-6581
Container-title:Physical Sciences Reviews
language:en
Short-container-title:

Author:

García Laura¹,Ruíz Ingrid²,Gómez José A.²

Affiliation:

1. Faculty of Chemistry, University of Havana , 10400 , Havana , Cuba

2. R&D Quality Control Department , Center of Molecular Immunology , 11300 , Havana , Cuba

Abstract

Abstract At the Center of Molecular Immunology (Havana, Cuba), the fusion protein SARS-CoV-2 S protein (RBD)-hFc was synthesized linking the receptor-binding domain (RBD) of the SARS-CoV-2 virus and the crystallizable fragment of a human immunoglobulin. This fusion protein was used in the construction of a diagnostic device for COVID-19 called UMELISA SARS-CoV-2-IgG. Given the relevance of this protein, the characterization of three batches (A1, A2 and A3) was carried out. The molecular weight of the protein was determined to be 120 kDa, using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Its isoelectric point was estimated between 8.3 and 9 by isoelectric focusing. The molecular integrity was evaluated by size exclusion liquid chromatography and SDS-PAGE after one year of the production of the protein; the presence of aggregates and fragments was detected. Batches A1 and A2 have a purity percentage higher than 95% and they can be used for the construction of new diagnostic devices.

Publisher

Walter de Gruyter GmbH

Subject

General Physics and Astronomy,General Materials Science,General Chemistry

Link

https://www.degruyter.com/document/doi/10.1515/psr-2021-0164/pdf

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