An efficient system to generate truncated human angiotensin converting enzyme 2 (hACE2) capable of binding RBD and spike protein of SARS-CoV2

Abstract

Human angiotensin converting enzyme 2 (hACE2) mediates the cell entry of both SARS-CoV and SARS-CoV2 and can be used as a drug target. The DNA encoding the truncated hACE2 (30-356aa) was cloned into pET-28a (+) and expressed in Escherichia coli Rosetta (DE3). The recombinant hACE2 (rhACE2) was purified by affinity chromatography on a Ni-NTA column and characterized with SDS-PAGE and Western blot. The binding activity of rhACE2 to Spike protein of SARS-CoV2 was evaluated in S protein-overexpressed HEK293A cells (HEK293A-SP cells) through flow cytometry. The prokaryotic expression system is able to produce approximately 75 mg protein per liter, which would be useful for infection mechanism study, and drug screening and development of SARS-CoV2.

Highly sensitive and selective surface plasmon resonance biosensor for the detection of SARS-CoV-2 spike S1 protein

“…1), and they are the major targets for SARS-CoV-2 therapy and detection. [39][40][41]52 Antigen detection is a good choice for testing viral loading in early stages of the COVID-19 disease. This work explored 2D MXene-based SPR sensing strategy for detecting SARS-CoV-2 spike S1 protein abbreviated as S1 protein.…”

Utilization of Receptor-Binding Domain of SARS-CoV-2 Spike Protein Expressed in Escherichia coli for the Development of Neutralizing Antibody Assay

“…This system can be modified easily and set up in any molecular biology laboratory, offering a significant advantage in managing COVID-19 in low-income countries where the cost of commercial test kits can be obstacle and local variants are likely to emerge. On another note, a few previous studies also attempted to use E. coli as an expression system for ACE2 [ 26 , 27 ]. It remains to be investigated if the combination of hACE2 and E-RBD produced from E. coli could be used for the development of a NAb detection assay.…”