tetano
Editor, Senior Moderator
Anal Chim Acta
. 2023 Jul 11;1264:341300.
doi: 10.1016/j.aca.2023.341300. Epub 2023 Apr 29. Biopanning of specific peptide for SARS-CoV-2 nucleocapsid protein and enzyme-linked immunosorbent assay-based antigen assay
Pengxin Ma[SUP] 1 [/SUP], Junchong Liu[SUP] 1 [/SUP], Shuang Pang[SUP] 1 [/SUP], Wenhao Zhou[SUP] 2 [/SUP], Haipeng Yu[SUP] 1 [/SUP], Mingyang Wang[SUP] 1 [/SUP], Tao Dong[SUP] 1 [/SUP], Yanbo Wang[SUP] 1 [/SUP], Qiqin Wang[SUP] 3 [/SUP], Aihua Liu[SUP] 4 [/SUP]
Affiliations
The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread worldwide which triggered serious public health issues. The search for rapid and accurate diagnosis, effective prevention, and treatment is urgent. The nucleocapsid protein (NP) of SARS-CoV-2 is one of the main structural proteins expressed and most abundant in the virus, and is considered a diagnostic marker for the accurate and sensitive detection of SARS-CoV-2. Herein, we report the screening of specific peptides from the pIII phage library that bind to SARS-CoV-2 NP. The phage monoclone expressing cyclic peptide N1 (peptide sequence, ACGTKPTKFC, with C&C bridged by disulfide bonding) specifically recognizes SARS-CoV-2 NP. Molecular docking studies reveal that the identified peptide is bound to the "pocket" region on the SARS-CoV-2 NP N-terminal domain mainly by forming a hydrogen bonding network and through hydrophobic interaction. Peptide N1 with the C-terminal linker was synthesized as the capture probe for SARS-CoV-2 NP in ELISA. The peptide-based ELISA was capable of assaying SARS-CoV-2 NP at concentrations as low as 61 pg/mL (∼1.2 pM). Furthermore, the as-proposed method could detect the SARS-CoV-2 virus at limits as low as 50 TCID[SUB]50[/SUB] (median tissue culture infective dose)/mL. This study demonstrates that selected peptides are powerful biomolecular tools for SARS-CoV-2 detection, providing a new and inexpensive method of rapidly screening infections as well as rapidly diagnosing coronavirus disease 2019 patients.
Keywords: ELISA; Phage display; SARS-COV-2 virus; SARS-CoV-2 NP; Specific peptide.
. 2023 Jul 11;1264:341300.
doi: 10.1016/j.aca.2023.341300. Epub 2023 Apr 29. Biopanning of specific peptide for SARS-CoV-2 nucleocapsid protein and enzyme-linked immunosorbent assay-based antigen assay
Pengxin Ma[SUP] 1 [/SUP], Junchong Liu[SUP] 1 [/SUP], Shuang Pang[SUP] 1 [/SUP], Wenhao Zhou[SUP] 2 [/SUP], Haipeng Yu[SUP] 1 [/SUP], Mingyang Wang[SUP] 1 [/SUP], Tao Dong[SUP] 1 [/SUP], Yanbo Wang[SUP] 1 [/SUP], Qiqin Wang[SUP] 3 [/SUP], Aihua Liu[SUP] 4 [/SUP]
Affiliations
- PMID: 37230729
- DOI: 10.1016/j.aca.2023.341300
The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread worldwide which triggered serious public health issues. The search for rapid and accurate diagnosis, effective prevention, and treatment is urgent. The nucleocapsid protein (NP) of SARS-CoV-2 is one of the main structural proteins expressed and most abundant in the virus, and is considered a diagnostic marker for the accurate and sensitive detection of SARS-CoV-2. Herein, we report the screening of specific peptides from the pIII phage library that bind to SARS-CoV-2 NP. The phage monoclone expressing cyclic peptide N1 (peptide sequence, ACGTKPTKFC, with C&C bridged by disulfide bonding) specifically recognizes SARS-CoV-2 NP. Molecular docking studies reveal that the identified peptide is bound to the "pocket" region on the SARS-CoV-2 NP N-terminal domain mainly by forming a hydrogen bonding network and through hydrophobic interaction. Peptide N1 with the C-terminal linker was synthesized as the capture probe for SARS-CoV-2 NP in ELISA. The peptide-based ELISA was capable of assaying SARS-CoV-2 NP at concentrations as low as 61 pg/mL (∼1.2 pM). Furthermore, the as-proposed method could detect the SARS-CoV-2 virus at limits as low as 50 TCID[SUB]50[/SUB] (median tissue culture infective dose)/mL. This study demonstrates that selected peptides are powerful biomolecular tools for SARS-CoV-2 detection, providing a new and inexpensive method of rapidly screening infections as well as rapidly diagnosing coronavirus disease 2019 patients.
Keywords: ELISA; Phage display; SARS-COV-2 virus; SARS-CoV-2 NP; Specific peptide.