Stable production of recombinant SARS-CoV-2 receptor-binding domain in mammalian cells with co-expression of a fluorescent reporter and its validation as antigenic target for COVID-19 serology testing

 

محفوظ في:
التفاصيل البيبلوغرافية
المؤلفون: Corrales Aguilar, Eugenia, Arias Arias, Jorge Luis, Molina Castro, Silvia Elena, Monturiol Gross, Laura, Lomonte, Bruno
التنسيق: artículo original
تاريخ النشر:2023
الوصف:SARS-CoV-2 receptor binding domain (RBD) recognizes the angiotensin converting enzyme 2 (ACE2) receptor in host cells that enables infection. Due to its antigenic specificity, RBD production is important for development of serological assays. Here we have established a system for stable RBD production in HEK 293T mammalian cells that simultaneously express the recombinant fluorescent protein dTomato, which enables kinetic monitoring of RBD expression by fluorescence microscopy. In addition, we have validated the use of this recombinant RBD in an ELISA assay for the detection of anti-RBD antibodies in serum samples of COVID-19 convalescent patients. Recombinant RBD generated using this approach can be useful for generation of antibody-based therapeutics against SARS-CoV-2, as well serological assays aimed to test antibody responses to this relevant virus.
البلد:Kérwá
المؤسسة:Universidad de Costa Rica
Repositorio:Kérwá
اللغة:Inglés
OAI Identifier:oai:kerwa.ucr.ac.cr:10669/88167.2
الوصول للمادة أونلاين:https://doi.org/10.1016/j.btre.2022.e00780
https://hdl.handle.net/10669/88167.2
كلمة مفتاحية:SARS-CoV-2
Recombinant RBD expression
Fluorescent protein
Mammalian cells
COVID-19