Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera

Lee, Hyojin; Kim, Eun-Ju; Song, Jae-Young; Choi, Jeong Soo; Lee, Ji Youn; Cho, In-Soo; Shin, Yeun-Kyung

Immunization with N protein for rabbit polyclonal antibody and cattle positive control sera production Animal experiments were conducted according to the protocol of the Institutional Animal Care and Use Committee of the Republic of Korea. Rabbit polyclonal antibody and mouse mAbs were generated after four immunizations with recombinant NP. Polyclonal antibodies were produced in rabbits after four injections of 500 g of the NP antigen mixed with adjuvant at two-week intervals. The NP-specific mAbs were synthesized as previously described [2] . Briefly, eight mice were injected four times with 100 g of recombinant NP. Following immunization, the mouse spleen cells were collected and fused with myeloma cells to generate mAbs. Among the mouse hybridoma cell lines, 14 clones were selected according to specific interactions with NP using indirect ELISA.