Monoclonal antibodies to newcastle disease virus: delineation of four epitopes on the HN glycoprotein

Abstract

Eighteen independent hybridomas producing monoclonal antibodies to Newcastle disease virus have been prepared by fusion of SP2 cells with spleen lymphocytes from a BALB/c mouse immunized with intact UV-inactivated Newcastle disease virus strain Australia-Victoria. They have been divided into three groups on the basis of radioimmunoprecipitation, infected cell surface and cytoplasmic fluorescence, and isotype. The anti-HN group is made up of nine antibodies which give surface fluorescence on infected cells and immunoprecipitate the HN glycoprotein. These antibodies bind to HN in nitrocellulose transfers of sodium dodecyl sulfate gels, but only if it has been neither reduced nor boiled. To varying degrees, all of these HN antibodies neutralize infectivity. These results suggest that they recognize exposed determinants of a conformational nature on the native HN molecule. They have been used in competition antibodybinding radioimmunoassays and additive neutralization assays, and on the basis of

Neutralization map of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus: domains recognized by monoclonal antibodies that prevent receptor recognition

“…Hybridomas and MAbs. The preparation of hybridomas and the initial characterization of MAbs have been described previously (12,13,18).…”

“…We initially characterized a panel of monoclonal antibodies (MAbs) to antigenic sites 1, 2, 3, and 4 on the surface of the HN glycoprotein of the AV isolate (13). However, many MAbs produced by hybridomas isolated in subsequent fusions were shown to map to sites which overlap two of the original four sites and were named to reflect this, hence sites 12, 14, and 23 (12,18).…”

The antiviral activity of antibodies in vitro and in vivo

“…Additive effects are also observed for antibodies binding to the paramyxovirus Newcastle disease virus (NDV). Binding of single mAbs against the NDV 210 PARREN AND BURTON hemagglutinin-neuraminidase (HN) glycoprotein leaves a large persistent fraction of nonneutralized virus (Iorio and Bratt, 1983). The persistent fraction appeared to be bound by neutralizing mAb, as it was almost completely neutralized by the addition of anti-antibody (Iorio and Bratt, 1984).…”

Package of NDV-Pseudotyped HIV-Luc Virus and Its Application in the Neutralization Assay for NDV Infection

“…For these neutralizing antibodies, the HI assay would be inadequate. More commonly, neutralizing antibodies against NDV are evaluated by VN tests with native NDV [56,57]. However, the VN test with native NDV is time-consuming, requiring at least 4 days, and less-objective.…”