Preparation and identification of monoclonal antibodies against σB protein of Muscovy duck reovirus.

σB protein of Muscovy duck reovirus(MDRV) was expressed in Escherichia coli BL21(DE3) harboring pET30-σB plasmid.Female BALB/c mice were immunized by using the purified σB protein as immunogen to screen monoclonal antibodies(mAbs) against the σB protein by using indirect enzyme linked immunosorbent assay,Western-blot,and immunohistochemistry assay.The results showed that four strains of hybridoma cell lines stably secreting mAbs against σB,designated as A3F,B2B,C3E,and D2G respectively,were obtained,and they belonged to subtype IgG2b,IgG2b,IgM,or IgM respectively.The four mAbs in the ascites of mice inoculated with the four hybridoma cell lines could react specifically with the σB protein expressed in MDRV-infected duck embryo fibroblasts,indicating that the mAbs could recognize the σB protein in natural conformation.