Razi Vaccine & Serum Research Institute Co-Publisher: Negah Scientific Publisher Archives of Razi Institute 0365-3439 80 1 2025 02 01 Unleashing Monoclonal Antibodies: Targeting Covid-19’s Nucleocapsid Protein and Spike Antigens طراحی و تولید آنتی بادی های مونوکلونال با هدف قرار دادن آنتی ژن های Nucleocapsid و Spike 75 84 132037 10.32592/ARI.2025.80.1.75 EN Rasool Madani Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran Fariba Golchinfar Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran Mahsa Hezarosi Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran Tara Emami Razi Vaccine and Serum Research Institute Journal Article 2024 05 04 Since the end of 2019, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected many people globally. Diagnosis and treatment of patients have a pivotal role in surviving them. Two units of virus namely, Nucleocapsid protein and Spike proteins play important roles in entering and affecting cells. These two substances can be good targets for producing monoclonal antibodies which can be useful in treatment, serological diagnosis tests, and even prevention by vaccination.<br />The Nucleocapsid protein and Spike proteins of SARS‐CoV‐2 were purchased by Razi Vaccine and Research Institue in Karaj in 2020. Then they were separately injected into mice with appropriate dosage of adjuvants. The spleen cells of immunized mice were fused with myeloma cells. The best antibody producer clones were opted for further evaluation tests. Western Blot and ELISA were used to demonstrate the immunoreactivity of the recombinant Np and S proteins. Finally, we freeze the best clones in nitrogen.<br />By using an ELISA test, eight clone antibodies namely, 3G1, 3G2, 3E7, H11, A11, F10, B11, and 2F6 were identified. These monoclonal antibodies were found to be against the S and N p antigens of SARS-CoV-2. Additionally, the results of the western blot test indicated that each of these antibodies had antigenic sites against Spike and Nucleocapsid protein independently, and the isotyping test revealed that they were from IgG (2a, 2b) or IgM class antibodies.<br />The development of monoclonal antibodies has the potential to help in both diagnosis and treatment. The Nucleocapsid protein and Spike protein from SARS-CoV-2 exhibit a great deal of promise for creating a new generation of monoclonal antibodies. Moreover, a thorough method for the early diagnosis of COVID-19 can be offered by combining the detection of Nucleocapsid protein and Spike proteins.. https://archrazi.areeo.ac.ir/article_132037_3aa68cfdf0a632e666c64641a807a0a8.pdf