Abstract
Antigen Used for Immunization
Recombinant SARS-CoV-2 nucleocapsid protein was used as an immunogen.
Method of Immunization
BALB/c mice (female) were subcutaneously injected with 250 μg recombinant SARS-CoV-2 nucleocapsid protein in Freund's complete adjuvant at the ratio (1:1) followed by Freund's incomplete adjuvant. Three booster injections were given with the same dose of antigen without adjuvant. (1) The titer of mouse serum was determined by enzyme-linked immunosorbent assay.1
Parental Cell Line Used for Fusion
SP2/0 mouse myeloma cells.
Selection and Cloning Procedure
The fused cells were cultured in hypoxanthine-aminopterin-thymidine medium, and the positive wells were subcloned three times by limiting dilution.
Heavy and Light Chains of Immunoglobulin
The two monoclonal antibodies were identified as immunoglobulin G1 isotype.
Specificity
Specificity was evaluated by western blotting analysis.
Specific Antigen Identified
SARS-CoV-2 nucleocapsid protein.
Availability
Author's Contribution
P.W.: Writing–review and editing.
Footnotes
Author Disclosure Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Funding Information
This work was supported by the Science and Technology Support Program of Guizhou Province (Grant No. [2020]4Y191).
Address correspondence to:
Pei Wei
Department of Immunology
Zhuhai Campus of Zunyi Medical University
No. 368 Golden Coast Road
Zhuhai, Guangdong 519040
China
E-mail: weipei@zmu.edu