Plasmid to Produce Recombinant SARS-CoV-2 Nucelocapsid Protein

Diagnostic Assays

At a Glance

​Researchers at Colorado State University have developed several plasmids and purification protocols for the expression and purification of a soluble SARS-CoV-2 nucleocapsid protein for use in diagnostic assays, in response to the SARS-CoV-2/COVID-19 outbreak.

Overview

SARS-CoV-2 Nucleocapsid Protein Expression Plasmid
pBG690 SARS-CoV-2 Nucleocapsid Protein Expression Plasmid

Highlights:

  • Bacterial expression plasmid encodes a bacterial codon-optimized fragment of the SARS-CoV-2 nucleocapsid protein
  • Plasmid was sequence verified
  • Bacterial expression protocol can produce monomeric nucleocapsid protein (as confirmed by mass spectroscopy) with greater than 95% purify
  • Purified nucleocapsid protein has been tested by collaborators for antigenicity in mice immunized against the recombinant protein as well as bats infected with SARS-CoV-2
  • Protein expressed and purified from this plasmid is antigenic
  • Protein can be used for ELISAs as well as other assays to indicate presence of SARS-CoV-2 nucleocapsid protein antibodies in patient samples
Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] Expression Plasmids

Highlights:

  • Recognizes SARS-CoV2 Nucleocapsid protein
  • mBG17 did show cross reaction with SARS-CoV nucleocapsid protein and minor cross-reaction with HuCoV-NL63 nucleocapsid protein was observed by ELISA,
  • no cross-reaction was observed with other coronavirus N proteins.
  • Suitable for ELISA, immunofluorescence assay, and western blot applications
​Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86] Expression Plasmids

Highlights:

  • Recognizes SARS-CoV2 Nucleocapsid protein
  • mBG86 showed no cross reaction with SARS-CoV nucleocapsid or any of the other coronavirus N proteins tested by ELISA
  • mBG86 is very specific for SARS-CoV-2 nucleocapsid protein.
  • Suitable for ELISA, immunofluorescence assay, and western blot applications

Benefits

  • Plasmid can be licensed for in-house production – allows for larger amounts of antigen for diagnostic testing
  • SARS-CoV-2 nucleoprotein is a major diagnostic antigen for detection of active infection and immunity

Applications

  • SARS-CoV-2 diagnostic assays
Last Updated: January 2024
Gloved scientist with COVID samples
Inventors
Reference Number
2020-090
Licensing Manager

Steve Foster
Steve.Foster@colostate.edu
970-491-7100