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Sandwich ELISA was carried out by coating 80027-1-RR at 80 ng/well followed by blocking and adding different concentration of eukaryotic expressed N protein (0.5-1000 ng/mL). HRP-conjugated80026-1-RR was used at 1 μg/mL for detection. Signal was developed with TMB and stopped by H2SO4 . Signal strength was measured by absorbance at 450 nm.
The nucleocapsid (N) protein has multiple functions including formation of nucleocapsids, signal transduction virus budding, RNA replication, and mRNA transcription. N protein is an important antigen for coronavirus, and it is normally highly conserved, with a molecular weight of about 50 kDa. it can be used as a marker in diagnostic assays due to its high immunogenicity (PMID: 32416961, PMID: 32235387). A sandwich ELISA for COVID-19 N Protein can be assembled by using 80027-1-RR as capture antibody and conjugated 80026-1-RR for detection.