Magnetic Bead-Based Immunoassay Allows Rapid, Inexpensive, and Quantitative Detection of Human SARS-CoV-2 Antibodies

Huergo, Luciano F.; Selim, Khaled A.; Conzentino, Marcelo S.; Gerhardt, Edileusa C. M.; Santos, Adrian R. S.; Wagner, Berenike; Alford, Janette T.; Deobald, Nelli; Pedrosa, Fabio O.; de Souza, Emanuel M.; Nogueira, Meri B.; Raboni, Sônia M.; Souto, Dênio; Rego, Fabiane G. M.; Zanette, Dalila L.; Aoki, Mateus N.; Nardin, Jeanine M.; Fornazari, Bruna; Morales, Hugo M. P.; Borges, Vânia A.; Nelde, Annika; Walz, Juliane S.; Becker, Matthias; Schneiderhan-Marra, Nicole; Rothbauer, Ulrich; Reis, Rodrigo A.; Forchhammer, Karl

[Image: see text] Immunological methods to detect SARS-CoV-2 seroconversion in humans are important to track COVID-19 cases and the humoral response to SARS-CoV-2 infections and immunization to future vaccines. The aim of this work was to develop a simple chromogenic magnetic bead-based immunoassay which allows rapid, inexpensive, and quantitative detection of human antibodies against SARS-CoV-2 in serum, plasma, or blood. Recombinant 6xHis-tagged SARS-CoV-2 Nucleocapsid protein was mobilized on the surface of Ni(2+) magnetic beads and challenged with serum or blood samples obtained from controls or COVID-19 cases. The beads were washed, incubated with anti-human IgG-HPR conjugate, and immersed into a solution containing a chromogenic HPR substrate. Bead transfer and homogenization between solutions was aided by a simple low-cost device. The method was validated by two independent laboratories, and the performance to detect SARS-CoV-2 seroconversion in humans was in the same range as obtained using the gold standard immunoassays ELISA and Luminex, though requiring only a fraction of consumables, instrumentation, time to deliver results, and volume of sample. Furthermore, the results obtained with the method described can be visually interpreted without compromising accuracy as demonstrated by validation at a point-of-care unit. The magnetic bead immunoassay throughput can be customized on demand and is readily adapted to be used with any other 6xHis tagged protein or peptide as antigen to track other diseases.