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The production of the N recombinant monoclonal antibody is carried out through a systematic and thorough process to ensure its quality and specificity. The journey begins by isolating B cells from the spleen of an immunized animal, where the recombinant human SARS-CoV-2 N protein (1-419aa) is utilized as the immunogen. The RNA is then extracted from these B cells and converted into cDNA through reverse transcription. Using specific primers designed for the antibody constant regions, the N antibody genes are amplified and inserted into an expression vector. The human IgG1 Fc is integrated into the vector, downstream of the N antibody gene. The biotin is also inserted into the vector. The recombinant vector is then introduced into host cells through transfection, enabling the production of the N recombinant monoclonal antibody. Following an appropriate incubation period, the antibody is collected from the cell culture supernatant and subjected to purification using affinity chromatography, ensuring a high level of purity. To validate its specificity and functionality in detecting human SARS-CoV-2 N protein, the purified N recombinant monoclonal antibody undergoes rigorous characterization through ELISA. This meticulous production process yields a reliable and effective N recombinant monoclonal antibody, serving as a valuable tool in various research related to SARS-CoV-2.
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