Antigenicity of Japanese encephalitis virus envelope glycoprotein V3 (E) and its cyanogen bromide cleaved fragments examined by monoclonal antibodies and Western blotting.

Purified Japanese encephalitis (JE) virus was solubilized under reducing condition by using 2-merceptoethanol (2 ME) or nonreducing condition without 2 ME and its structural proteins were separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE), followed by the Western blotting using polyclonal and monoclonal antibodies against JE. The mobilities and reactivities against polyclonal antiserum of V3 (E) and V2 (C) were reduced when virion was disrupted under reducing condition. The 54 K band corresponding to V3 was treated with cyanogen bromide (CNBr) and analyzed by the second SDS-PAGE and Western blotting.