Preparation And Identification Of The Monoclonal Antibody Against Newcastle Disease Virus

A hybridoma cell strain which can secrete monoclonal antibody (McAb) against Newcastle Disease viruses (NDV) was generated by using lymphocyte cell fusion technology. The McAb was developed and it's characteristics was identified. In the study, the Newcastle Disease viruses were replicated through virus inoculation in embryonated eggs. The NDV particles were purified by differential centrifugation. Then, Balb/C mice were immunized with the purified NDV particles which were fully emulsified with Freund's adjuvant. The spleen cells of immunized Balb/C mice were fused with NS0 mouse plasmacytoma cells using PEG-1500. The hybridoma cell secreting McAb against NDV were screened by indirect ELISA and the hemagglutination inhibition (HI) assay. Positive clones were sub-cloned by limited dilution. The characteristics of the McAb were identified by ELISA, HI and western-blotting. The McAb ELISA titers from cell culture supernatant and ascites were 1:160 and 1:3.2×104, respectively. The McAb showed no cross-reactivity to negative allantoic fluid and other avian virus antigens including AIV, IBV and IBDV. The HI assay had proved that the McAb had no HI activity. Western-blotting analysis showed that the McAb specifically reacted with the peptides of NDV. During four months, the hybirdoma cell can stably secrete McAb of NDV after serial passages and three times freezing and revival. The preparation of the McAb of NDV can provide a foundation for the virus research and a basis for development rapid diagnostic methods.