expression vectors containing the constant regions of the human igg1  (GenScript corporation)

Journal: Virology Journal

Article Title: Identification and application of a pair of noncompeting monoclonal antibodies broadly binding to the nucleocapsid proteins of SARS-CoV-2 variants including Omicron

doi: 10.1186/s12985-022-01827-w

Figure Lengend Snippet: The application of P301-F7 and P301-H5 in the detection of NPs in multiple biochemical assays. a Western blot analysis of the binding activities of P301-F7 and P301-H5 to NPs of SARS-CoV-2, SARS-CoV, and MERS-CoV, respectively. The His-tag at C-terminus of each NP detected by the His-specific antibody is used here as a positive control in the assay. b Flow cytometry analysis of the binding activities of P301-F7 and P301-H5 to the SARS-CoV-2 and SARS-CoV NPs. Commercial polyclonal antibodies of one kind are used here as a positive control. c Immunofluorescence analysis of the inhibition effect of SARS-CoV-2 live virus by Remdesivir. Intracellular expression of NP was detected by staining Vero E6 cells infected with SARS-CoV-2 by P301-F7. The nuclei is stained with Hoechst. d The immune spot assay analysis of P301-F7 for recognizing the Vero E6 cells infected with SARS-CoV-2 live virus represented by blue spots. e The immune spot assay analysis of the inhibition efficiency of SARS-CoV-2 live virus in Vero E6 cells by P2C-1F11. IgG1 is a negative control of irrelevant antibody. VC: virus control without antibody. CC: cell control without virus and antibody. Each experiment was performed twice and one representative result was shown

Article Snippet: Then heavy chain and light chain genes were separately amplified, sequenced, synthesized, and cloned into the backbone of antibody expression vectors containing the constant regions of human IgG1 (Sangon Biotech).

Techniques: Western Blot, Binding Assay, Positive Control, Flow Cytometry, Immunofluorescence, Inhibition, Virus, Expressing, Staining, Infection, Spot Test, Negative Control, Control