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.
KW - COVID-19
KW - diagnostics
KW - ELISA
KW - peptide epitopes
KW - SARS-CoV-2
UR - http://www.scopus.com/inward/record.url?scp=85171863716&partnerID=8YFLogxK
U2 - 10.1021/acsbiomedchemau.3c00010
DO - 10.1021/acsbiomedchemau.3c00010
M3 - Article
C2 - 38404747
AN - SCOPUS:85171863716
SN - 2694-2437
VL - 4
SP - 37
EP - 52
JO - ACS Bio and Med Chem Au
JF - ACS Bio and Med Chem Au
IS - 1
ER -
