The Preparation And Application Of Monoclonal Antibodies Against Human Adenovirus Type3and7

[Objective] To prepare and identify monoclonal antibodies (MAbs) from miceimmunized with human adenovirus type3and type7-chimeric adenovirus.[Methods] A chimeric adenovirus designated “rAdMHE3”, which was constructed byreplacing hexon hypervariable region5(HVR5) of an human adenovirus type3(HAdv-3)-based vector designated “Ad3EGFP” with HVR5of HAdv-7, was used toimmunized BALB/c mice. Using hybridoma technique, the MAbs against rAdMHE3was prepared and screened by indirect enzyme-linked immunosorbent assays(ELISAs). The neutralizing MAbs against Ad3EGFP or HAdv-7were then screenedusing adenovirus neutralization assays in vitro， and identified using immunoblottingassays, indirect ELISAs, and antibody-binding-competition assays. The MAbs whichcan detect HAdv virions from cell culture were screened using directimmunofluorescence assays (IFAs).[Results] Among the twenty-two MAbs against rAdMHE3obtained, four showedneutralizing activity against HAdv-7and three against Ad3EGFP. Furtheridentification determined that the four HAdv-7-neutralizing MAbs recognize theneutralizing site within HVR5of HAdv-7and the three Ad3EGFP-neutralizingrecognize HVR4of Ad3EGFP. Using direct IFAs, four MAbs could detect HAdv-7 and two MAbs could detect HAdv-3and ten MAbs could detect HAdv-3,4, and7.[Conclusion] This study provides preliminary confirmation that the strategy, whichinvolves the incorporation of antigen into HVRs of an HAdv-3-based vector, could befeasibly developed to generate MAbs. HVR4of HAdv-3may contain a linearserotype-specific neutralizing epitope.