Antibodies Produced Toward Recombinant RBD and Nucleocapsid Neutralize SARS-COV-2

Area of Science:

• Virology
• Immunology
• Biotechnology

Background:

• SARS-CoV-2 caused a global pandemic, infecting millions.
• The Spike protein's Receptor Binding Domain (RBD) is key for host cell entry.
• The N protein is essential for viral genome packaging.

Purpose of the Study:

• To develop recombinant SARS-CoV-2 proteins for potential vaccine development.
• To assess the immunogenicity and neutralizing capacity of RBD and N proteins.

Main Methods:

• Bioinformatic approaches including codon adaptation and protein stability.
• Cloning, expression, and purification of recombinant RBD and N proteins in E. coli.
• Animal immunization and antibody titer determination using ELISA and neutralization tests.

Main Results:

• Successfully expressed and purified RBD (28 kDa) and N (45 kDa) proteins, confirmed by SDS-PAGE and Western blotting.
• ELISA showed high antibody titers (up to 1:64000) in immunized mice sera.
• Neutralization tests confirmed the antibodies' ability to bind and neutralize SARS-CoV-2 proteins.

Conclusions:

• Recombinant RBD and N proteins effectively stimulate an immune response.
• The generated antibodies demonstrate in vitro virus-neutralizing potential.