The preparation and preliminary application of the group-specific monoclonal antibodies against the nonstructural glycoprotein 1 of dengue virus

Objective Prepare the group specific monoclonal antibodies(McAbs) against DENV NS1 antigens,to establish a double antibody sandwich ELISA which could detect the NS1 antigen of DENV 1～4.Methods BABL /c mice were immunized with recombinant DENV2-NS1 protein,which is expressed in Pichia Pastoris.Then,the splenocyte were collected and fused with mouse myeloma cell SP2 /0 by PEG.After the indirect ELISA screening and sub-cloning,hybridoma cell lines which could stably secrete group specific anti-NS1 monoclonal antibodies were prepared.A double antibody sandwich ELISA to detect the DENV 1 ～4 NS1 protein was established..Results Plenty of purified recombinant DENV2-NS1 proteins were expressed and 2 hybridoma cell lines,2D7B6B4 and 2D10E2F6 were obtained after traditional hybridomas technology.It was detected that both of them could co-react with DENV 1～4 and its NS1s.The titer which was detected by indirect ELISA was 1:8 000-1:16 000.The subtype of immunoglobulin was IgG2a.An early rapid testing method was initially established.Conclusion Successfully prepared two cross-serotype monoclonal antibodies that co-reactive with DENV 1～4 and the NS1s.A double antibody sandwich ELISA was established,making the great foundation of early rapid diagnosis to dengue fever.