Delivery of Antibody-Like Molecules, Monobodies, Capable of Binding with SARS-CoV-2 Virus Nucleocapsid Protein, into Target Cells

Autor:	Y V, Khramtsov, A V, Ulasov, T N, Lupanova, G P, Georgiev, A S, Sobolev
Rok vydání:	2022
Předmět:	HEK293 Cells SARS-CoV-2 Escherichia coli Biophysics Humans COVID-19 General Chemistry General Medicine Nucleocapsid Proteins Antibodies Viral Biochemistry
Zdroj:	Doklady Biochemistry and Biophysics. 506:220-222
ISSN:	1608-3091 1607-6729
DOI:	10.1134/s1607672922050088
Popis:	Based on previous studies, two antibody-like molecules, monobodies, capable of high-affinity interaction with the SARS-CoV-2 nucleocapsid protein (dissociation constant of tens of nM) were selected. For delivery to target cells, genetically engineered constructs containing monobody and TAT peptide, placed either at the N- or C-terminus of the resulting polypeptide, were produced and expressed in E. coli. The construct with the highest affinity to the SARS-CoV-2 nucleocapsid protein was revealed with the use of thermophoresis technique. Cellular thermal shift assay demonstrated the ability of this construct to interact with the nucleocapsid protein within HEK293T cells transfected with the SARS-CoV-2 nucleocapsid protein fused to the mRuby3 fluorescent protein. Replacement of TAT peptide to S10 shuttle peptide, containing endosomolytic peptide, significantly improved the penetration of the construct into the target cells.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d247044a897ecb58b13033df8a884ca8 https://doi.org/10.1134/s1607672922050088 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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