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The creation of the N recombinant monoclonal antibody follows a rigorous process to ensure its quality and specificity. It begins with the isolation of B cells from the spleen of an immunized animal, using the recombinant human SARS-CoV-2 N protein (1-419aa) as the immunogen. The RNA is extracted from these B cells and converted into cDNA through reverse transcription. The N antibody genes are then amplified using specific primers targeting the antibody constant regions and inserted into an expression vector. The human IgG1 Fc is integrated into the vector, downstream of the N antibody gene and the FITC is also inserted into the vector. This recombinant vector is transfected into host cells, allowing for the production of the N recombinant monoclonal antibody. The antibody is harvested from the cell culture supernatant and subjected to purification using affinity chromatography, resulting in a highly purified form. The purified N recombinant monoclonal antibody can be used to detect human SARS-CoV-2 N protein. Through this meticulous production process, a reliable and effective N recombinant monoclonal antibody is obtained, which serves as a valuable tool in various research related to SARS-CoV-2.
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