Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay

Waweru, G; Nyakundi, R; Kutima, B; Owuor, S; Konyino, G; Gitonga, J; Lugano, D; Maina, A; Musyoki, J; Ochola, L; Omondi, M; Kariuki, CK; Ogongo, P; Mwachari, C; Shee, F; Agoti, C; Sande, C; Uyoga, S; Kagucia, E; Agweyu, A; Bejon, P; Scott, JAG; Warimwe, GM; Ochola-Oyier, LI

COVID-19 Collection

Journal article

Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay

Abstract:: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continues to evolve, with mutations leading to the emergence of new variants. JN.1, a subvariant of omicron BA.2.86, has demonstrated marked immune escape and is now included in updated vaccine formulations. While reduced sensitivity has been reported for antibody assays using ancestral spike protein subunits to detect omicron-induced responses, the performance of full-length spike-based assays against omicron sublineages remains unclear. We aimed to compare the sensitivity of ELISA and Luminex assays using full-length spike proteins from the ancestral Wuhan strain and the JN.1 variant.MethodsWuhan and JN.1 full-length spike protein constructs were designed and expressed in Expi293F mammalian cells. In-house ELISAs based on previously validated protocols were used to measure anti-spike IgG levels. Additionally, a Luminex-based assay for anti-spike antibody detection was developed and validated. Both assays were applied to the following sample groups: pre-pandemic samples (designated "gold standard negatives"); PCR confirmed 2020 positives ("gold standard wildtype positives"); PCR confirmed 2024 positives ("gold standard omicron positives"); 2022 vaccinated individuals with verbal confirmed infection ("gold standard hybrid positives"); and 2024 household samples ("unknowns").ResultsWuhan spike protein showed a sensitivity of 100% (95% CI: 0.88-1.0) in detecting omicron-specific antibodies using gold standard omicron positives with JN.1 spike protein as a reference assay. Overall, across all samples, in ELISA, the Wuhan antigen had a sensitivity of 0.93 (95% CI: 0.89-0.95) and a specificity of 0.98 (95% CI: 0.94-0.99). The JN.1 antigen showed a sensitivity of 0.91 (95% CI: 0.87-0.94) and a specificity of 0.97 (95% CI: 0.93-0.99). In Luminex, sensitivity was 0.95 (95% CI: 0.91-0.97) for Wuhan and 0.94 (95% CI: 0.91-0.96) for JN.1. Specificity for both antigens in Luminex was 0.98 (95% CI: 0.94-0.99).ConclusionsBoth ELISA and Luminex assays showed comparable sensitivity and specificity for both Wuhan and JN.1 antigens, indicating that mutations in the JN.1 variant do not significantly impact assay performance. This suggests preserved antigenic recognition across variants.

Publication status:: Published

Peer review status:: Peer reviewed

Actions

Email

Email this record

Send the bibliographic details of this record to your email address.

Your Email
Please enter the email address that the record information will be sent to.

-
Your message (optional)
Please add any additional information to be included within the email.
Share
Cite

Cite this record

APA Style

Waweru, G., Nyakundi, R., Kutima, B., Owuor, S., Konyino, G., Gitonga, J., Lugano, D., Maina, A., Musyoki, J., Ochola, L., Omondi, M., Kariuki, C. K., Ogongo, P., Mwachari, C., Shee, F., Agoti, C., Sande, C., Uyoga, S., Kagucia, E., … Ochola-Oyier, L. I. (2025). Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay. Viruses, 17(9), 1248.

MLA Style

Waweru, G, et al. “Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay.” Viruses, vol. 17, no. 9, 2025, p. 1248.

Chicago Style

Waweru, G, R Nyakundi, B Kutima, et al. 2025. “Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay.” Viruses 17 (9): 1248.
Print