Mapping of functional domains on the influenza A virus RNA polymerase PB2 molecule using monoclonal antibodies

Abstract

Monoclonal antibodies against the PB2 of A/Puerto Rico/8/34 (A/PR/8/34) (H1N1) were prepared in order to define the functional domains of the RNA polymerase of influenza virus. The fifteen monoclonal antibodies that were generated were divided into 4 groups on the basis of ELISA binding to PB2 or its peptide fragments. Six Group I antibodies that bound to the PB2 N-terminal region (amino acids 1-104) did not inhibit transcription by the viral ribonucleoprotein complex. A single Group II antibody recognizing the region of amino acids 206-259 inhibited ApG-primed transcription. Groups III and IV antibodies bound to the C-terminal region of amino acids 660-759. Of these, Group III antibodies inhibited transcription. The present results identify multiple monoclonal antibody binding domains in PB2, two of which, when bound by antibodies, negatively affect viral RNA transcription.