tetano
Editor, Senior Moderator
Bioorg Chem
. 2021 Sep 23;116:105362.
doi: 10.1016/j.bioorg.2021.105362. Online ahead of print.
A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors
Xinjian Yin[SUP] 1 [/SUP], Litong Chen[SUP] 1 [/SUP], Siwen Yuan[SUP] 1 [/SUP], Lan Liu[SUP] 2 [/SUP], Zhizeng Gao[SUP] 3 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC[SUB]50[/SUB] of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC[SUB]50[/SUB] (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z' factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.
. 2021 Sep 23;116:105362.
doi: 10.1016/j.bioorg.2021.105362. Online ahead of print.
A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors
Xinjian Yin[SUP] 1 [/SUP], Litong Chen[SUP] 1 [/SUP], Siwen Yuan[SUP] 1 [/SUP], Lan Liu[SUP] 2 [/SUP], Zhizeng Gao[SUP] 3 [/SUP]
Affiliations
- PMID: 34598089
- PMCID: PMC8457910
- DOI: 10.1016/j.bioorg.2021.105362
Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC[SUB]50[/SUB] of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC[SUB]50[/SUB] (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z' factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.