Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Ali H. Aldoukhi; Panayiotis Bilalis; Dana M. Alhattab; Alexander U. Valle-Pérez; Hepi H. Susapto; Rosario Pérez-Pedroza; Emiliano Backhoff-García; Sarah M. Alsawaf; Salwa Alshehri; Hattan Boshah; Abdulelah A. Alrashoudi; Waleed A. Aljabr; Manal Alaamery; May Alrashed; Rana M. Hasanato; Raed A. Farzan; Roua A. Alsubki; Manola Moretti; Malak S. Abedalthagafi; Charlotte A.E. Hauser

doi:10.1021/acsbiomedchemau.3c00010

Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Ali H. Aldoukhi, Panayiotis Bilalis, Dana M. Alhattab, Alexander U. Valle-Pérez, Hepi H. Susapto, Rosario Pérez-Pedroza, Emiliano Backhoff-García, Sarah M. Alsawaf, Salwa Alshehri, Hattan Boshah, Abdulelah A. Alrashoudi, Waleed A. Aljabr, Manal Alaamery, May Alrashed, Rana M. Hasanato, Raed A. Farzan, Roua A. Alsubki, Manola Moretti, Malak S. Abedalthagafi, Charlotte A.E. Hauser^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

Original language	English (US)
Pages (from-to)	37-52
Number of pages	16
Journal	ACS Bio and Med Chem Au
Volume	4
Issue number	1
DOIs	https://doi.org/10.1021/acsbiomedchemau.3c00010
State	Accepted/In press - 2023

Bibliographical note

Funding Information:
This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19). The authors acknowledge Mauricio Andres Aguilar Aguila Isaias for his support in creating the graphical abstract.

Funding Information:
This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19).

Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society

Keywords

COVID-19
diagnostics
ELISA
peptide epitopes
SARS-CoV-2

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Pharmaceutical Science
Drug Discovery

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1021/acsbiomedchemau.3c00010

Cite this

Aldoukhi, A. H., Bilalis, P., Alhattab, D. M., Valle-Pérez, A. U., Susapto, H. H., Pérez-Pedroza, R., Backhoff-García, E., Alsawaf, S. M., Alshehri, S., Boshah, H., Alrashoudi, A. A., Aljabr, W. A., Alaamery, M., Alrashed, M., Hasanato, R. M., Farzan, R. A., Alsubki, R. A., Moretti, M., Abedalthagafi, M. S., & Hauser, C. A. E. (Accepted/In press). Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection. ACS Bio and Med Chem Au, 4(1), 37-52. https://doi.org/10.1021/acsbiomedchemau.3c00010

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abstract = "The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.",

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author = "Aldoukhi, {Ali H.} and Panayiotis Bilalis and Alhattab, {Dana M.} and Valle-P{\'e}rez, {Alexander U.} and Susapto, {Hepi H.} and Rosario P{\'e}rez-Pedroza and Emiliano Backhoff-Garc{\'i}a and Alsawaf, {Sarah M.} and Salwa Alshehri and Hattan Boshah and Alrashoudi, {Abdulelah A.} and Aljabr, {Waleed A.} and Manal Alaamery and May Alrashed and Hasanato, {Rana M.} and Farzan, {Raed A.} and Alsubki, {Roua A.} and Manola Moretti and Abedalthagafi, {Malak S.} and Hauser, {Charlotte A.E.}",

note = "Funding Information: This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19). The authors acknowledge Mauricio Andres Aguilar Aguila Isaias for his support in creating the graphical abstract. Funding Information: This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19). Publisher Copyright: {\textcopyright} 2023 The Authors. Published by American Chemical Society",

year = "2023",

doi = "10.1021/acsbiomedchemau.3c00010",

language = "English (US)",

volume = "4",

pages = "37--52",

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Aldoukhi, AH , Bilalis, P , Alhattab, DM , Valle-Pérez, AU, Susapto, HH, Pérez-Pedroza, R, Backhoff-García, E, Alsawaf, SM, Alshehri, S, Boshah, H , Alrashoudi, AA, Aljabr, WA, Alaamery, M, Alrashed, M, Hasanato, RM, Farzan, RA, Alsubki, RA, Moretti, M, Abedalthagafi, MS & Hauser, CAE 2023, 'Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection', ACS Bio and Med Chem Au, vol. 4, no. 1, pp. 37-52. https://doi.org/10.1021/acsbiomedchemau.3c00010

TY - JOUR

T1 - Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

AU - Aldoukhi, Ali H.

AU - Bilalis, Panayiotis

AU - Alhattab, Dana M.

AU - Valle-Pérez, Alexander U.

AU - Susapto, Hepi H.

AU - Pérez-Pedroza, Rosario

AU - Backhoff-García, Emiliano

AU - Alsawaf, Sarah M.

AU - Alshehri, Salwa

AU - Boshah, Hattan

AU - Alrashoudi, Abdulelah A.

AU - Aljabr, Waleed A.

AU - Alaamery, Manal

AU - Alrashed, May

AU - Hasanato, Rana M.

AU - Farzan, Raed A.

AU - Alsubki, Roua A.

AU - Moretti, Manola

AU - Abedalthagafi, Malak S.

AU - Hauser, Charlotte A.E.

N1 - Funding Information: This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19). The authors acknowledge Mauricio Andres Aguilar Aguila Isaias for his support in creating the graphical abstract. Funding Information: This work was financially supported by King Abdullah University of Science and Technology (KAUST) and by King Abdulaziz City for Science and Technology (KACST) with a funded grant (4419-KACST COVID-19). Publisher Copyright: © 2023 The Authors. Published by American Chemical Society

PY - 2023

Y1 - 2023

N2 - The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

AB - The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

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Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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