Dimeric ACE2-FC Is Equivalent to Monomeric ACE2 in the Surrogate Virus Neutralization Test

Kolesov, D.E.; Gaiamova, E.A.; Orlova, N.A.; Vorobiev, I.I.

Авторы: Kolesov Denis E., Gaiamova Elizaveta A., Orlova Nadezhda A., Vorobiev Ivan I.
Журнал: Biochemistry (Moscow)
Том: 88
Номер: 9
Год издания: 2023
Издательство: Pleiades Publishing, Ltd
Местоположение издательства: Road Town, United Kingdom
Первая страница: 1274
Последняя страница: 1283
DOI: 10.1134/s0006297923090079
Аннотация: Angiotensin-converting enzyme 2 (ACE2) is the main cellular receptor for the dangerous sarbecovirusesSARS-CoV and SARS-CoV-2. Its recombinant extracellular domain is used to monitor the level of protective humoralimmune response to a viral infection or vaccine using the surrogate virus neutralization test (sVNT). Soluble ACE2 isalso considered as an option for antiviral therapy potentially insensitive to the changes in the SARS-CoV-2 spike protein.Extensive testing of the samples of patient’s serum by the sVNT method requires using preparations of ACE2 or ACE2conjugates with constant properties. We have previously obtained a cell line that is a producer of a soluble monomericACE2 and showed that this ACE2 variant can be used in sVNT, preferably as a conjugate with horseradish peroxidase.A cell line that generates an ACE2-Fc fusion protein with high productivity, more than 150 mg/liter of the target proteinwhen cultured in a stirred flask, was obtained for producing a stable and universally applicable form of soluble ACE2.The affinity-purified ACE2-Fc fusion contains a mixture of dimeric and tetrameric forms, but allows obtaining linear responsecurves for inhibition of binding with the receptor-binding domain of the SARS-CoV-2 spike protein by antibodies.The ACE2-Fc-HRP-based sVNT testing system can be used for practical measurements of the levels of virus-neutralizingantibodies against various circulating variants of the SARS-CoV-2 virus.
Добавил в систему: Воробьев Иван Иванович

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Dimeric ACE2-FC Is Equivalent to Monomeric ACE2 in the Surrogate Virus Neutralization Testстатья

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