Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2

Kyosei, Yuta; Namba, Mayuri; Yamura, Sou; Takeuchi, Rikiya; Aoki, Noriko; Nakaishi, Kazunari; Watabe, Shigeo; Ito, Etsuro

doi:10.3390/diagnostics10080594

Cited by 45 publications

(54 citation statements)

References 25 publications

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“…Adapting peptide 1 to a chemiluminescence enzyme immunoassay or a similar assay could improve its sensitivity for detection of SARS-CoV-2. 8 , 39 Because of its selectivity in biological media, peptide 1 could be utilized for the direct detection of SARS-CoV-2 in bodily fluids. Improved diagnostics are a topic of intense COVID-19 research as they may provide rapid, reliable, and early detection.…”

Section: Discussionmentioning

confidence: 99%

De Novo Discovery of High-Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

et al. 2020

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The β-coronavirus SARS-CoV-2 has caused a global pandemic. Affinity reagents targeting the SARS-CoV-2 spike protein are of interest for the development of therapeutics and diagnostics. We used affinity selection–mass spectrometry for the rapid discovery of synthetic high-affinity peptide binders for the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. From library screening with 800 million synthetic peptides, we identified three sequences with nanomolar affinities (dissociation constants K d = 80–970 nM) for RBD and selectivity over human serum proteins. Nanomolar RBD concentrations in a biological matrix could be detected using the biotinylated lead peptide in ELISA format. These peptides do not compete for ACE2 binding, and their site of interaction on the SARS-CoV-2-spike-RBD might be unrelated to the ACE2 binding site, making them potential orthogonal reagents for sandwich immunoassays. These findings serve as a starting point for the development of SARS-CoV-2 diagnostics or conjugates for virus-directed delivery of therapeutics.

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Section: Discussionmentioning

confidence: 99%

De Novo Discovery of High-Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

et al. 2020

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“…Adapting peptide 1 to a chemiluminescence enzyme immunoassay or a similar assay could improve its sensitivity for detection of SARS-CoV-2. 8,37 Because of its selectivity in biological media, peptide 1 could be utilized for the direct detection of SARS-CoV-2 in bodily fluids. Improved diagnostics are a topic of intense COVID-19 research as they may provide rapid, reliable and early detection.…”

Section: Discussionmentioning

confidence: 99%

De Novo Discovery of High Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

Pomplun

Jbara

Quartararo

et al. 2020

Preprint

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The β-coronavirus SARS-CoV-2 has caused a global pandemic. Affinity reagents targeting the SARS-CoV-2 spike protein, the most exposed surface structure of the virus, are of interest for the development of therapeutics and diagnostics. We used affinity selection-mass spectrometry for the rapid discovery of synthetic high affinity peptide binders for the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. From library screening with 800 million synthetic peptides, we identified three sequences with nanomolar affinities (dissociation constants Kd = 80 to 970 nM) for RBD and selectivity over human serum proteins. Picomolar RBD concentrations in biological matrix could be detected using the biotinylated lead peptide in ELISA format. These peptides might associate with the SARS-CoV-2-spike-RBD at a site unrelated to ACE2 binding, making them potential orthogonal reagents for sandwich immunoassays. We envision our discovery as a robust starting point for the development of SARS-CoV-2 diagnostics or conjugates for virus directed delivery of therapeutics.Abstract Figure

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“… Ultrasensitive ELISA coupled with Thio-NAD cycling for SARS-CoV-2. Reprinted with permission from ref ( 80 ). Copyright (2020) MDPI (Basel, Switzerland).…”

Section: Current Diagnostic Methods For Covid-19mentioning

confidence: 99%

COVID-19 Antibody Tests and Their Limitations

2021

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COVID-19, caused by the SARS-CoV-2 virus, has developed into a global health crisis, causing over 2 million deaths and changing people’s daily life the world over. Current main-stream diagnostic methods in the laboratory include nucleic acid PCR tests and direct viral antigen tests for detecting active infections, and indirect human antibody tests specific to SARS-CoV-2 to detect prior exposure. In this Perspective, we briefly describe the PCR and antigen tests and then focus mainly on existing antibody tests and their limitations including inaccuracies and possible causes of unreliability. False negatives in antibody immunoassays can arise from assay formats, selection of viral antigens and antibody types, diagnostic testing windows, individual variance, and fluctuation in antibody levels. Reasons for false positives in antibody immunoassays mainly involve antibody cross-reactivity from other viruses, as well as autoimmune disease. The spectrum bias has an effect on both the false negatives and false positives. For assay developers, not only improvement of assay formats but also selection of viral antigens and isotopes of human antibodies need to be carefully considered to improve sensitivity and specificity. For clinicians, the factors influencing the accuracy of assays must be kept in mind to test patients using currently imperfect but available tests with smart tactics and realistic interpretation of the test results.

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Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2

Cited by 45 publications

References 25 publications

De Novo Discovery of High-Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

De Novo Discovery of High-Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

De Novo Discovery of High Affinity Peptide Binders for the SARS-CoV-2 Spike Protein

COVID-19 Antibody Tests and Their Limitations

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