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Sandwich ELISA was carried out by coating 67666-1-Ig at 80 ng/well followed by blocking and adding different concentration of eukaryotic expressed N protein (0.5-1000 ng/mL). HRP-conjugated clone 67666-2-Ig 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).67666-1-Ig can be used as capture antibody. 67666-2-Ig can be used as detection antibody.