The ongoing emergence of severe acute respiratory syndrome caused by the new coronavirus (SARS-CoV-2) variants requires swift actions in identifying specific antigens and optimizing vaccine development to maximize the humoral response of the patient. Measuring the specificity and the amount of antibody produced by the host immune system with high throughput and accuracy is critical to develop timely diagnostics and therapeutic strategies. Motivated by finding an easy-to-use and cost-effective alternative to existing serological methodologies for multiplex analysis, we develop a proof-of-concept multiplex nanoplasmonic biosensor to capture the humoral response in serums against multiple antigens. Nanoplasmonic sensing relies on the wavelength shift of the localized surface plasmon resonance (LSPR) peak of gold nanostructures upon binding interactions between the antibodies and the immobilized antigens. Here the antigens are first immobilized on different sensing areas by using a mono-biotinylation system based on the high affinity interaction between biotin and streptavidin. We then validate the multiplex platform by detecting the presence of 3 monoclonal antibodies against 3 antigens (2 different hemagglutinins (HAs) from influenza viruses, and the SARS-CoV-2 Spike RBD (receptor binding domain)). We also measure the humoral response in murine sera collected before and after its immunization with the SARS-CoV-2 Spike protein, in good agreement with the results obtained by the ELISA assay. Our nanoplasmonic assays have successfully demonstrated multiple serum antibody profiling, which can be further integrated with microfluidics as an effective high throughput screening platform in future studies for the ongoing SARS-CoV-2 vaccine development.
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