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Soluble SARS-CoV-2 RBD and human ACE2 peptidase domain produced in Drosophila S2 cells show functions evoking virus-cell interface

Protein Sci. 2023-08; 
Federico Carrión, Florencia Rammauro, Natalia Olivero-Deibe, Martín Fló, María Magdalena Portela, Analía Lima, Rosario Durán, Otto Pritsch, Sergio Bianchi
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Gene Synthesis … a STOP codon, were optimized for Drosophila codon usage and synthesized by Genscript. … Strep-tag was removed by incubating overnight with Enterokinase-His (Genscript) (20 U mg-1)… Get A Quote

Abstract

The interaction between the receptor-binding domain (RBD) of the spike glycoprotein of SARS-CoV-2 and the peptidase domain of the human angiotensin-converting enzyme 2 (ACE2) allows the first specific contact at the virus-cell interface making it the main target of neutralizing antibodies. Here, we show a unique and cost-effective protocol using Drosophila S2 cells to produce both RBD and soluble human ACE2 peptidase domain (shACE2) as thermostable proteins, purified via Strep-tag with yields >40 mg L in a laboratory scale. Furthermore, we demonstrate its binding with K values in the lower nanomolar range (independently of Strep-tag removal) and its capability to be blocked by serum antibodies in a competit... More

Keywords

ACE2, Drosophila, RBD, S2 cells, SARS-CoV-2, binding, microheterogeneity, neutralization, virus-cell interface