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Identification of L2 Open Reading Frame Gene Products of Bovine Papillomavirus Type 1 Using Monoclonal Antibodies

¹ Department of Pathology, Georgetown University Schools of Medicine and Dentistry, Washington, D.C. 20007
² Laboratory of Tumor Virus Biology, Division of Cancer Etiology, National Cancer Institute, National Institutes of Health, U.S. Department of Health and Human Services, Building 41, Room D507, Bethesda, Maryland 20892
and³ Molecular Genetics Inc., Minnetonka, Minnesota 55343, U.S.A.

Four hybridoma cell lines producing monoclonal antibodies (MAbs) to bovine papillomavirus type 1 (BPV-1) L2 open reading frame (ORF) gene products have been established from mice immunized with a BPV-1 L2--galactosidase fusion protein. Hybridomas were selected and cloned (from over 700 hybridomas) on the basis of specific reactivity of supernatant fluids with BPV-1 L2 epitopes on disrupted BPV-1 particles and L2--galactosidase fusion proteins by ELISA and Western blotting, and with acetone-fixed frozen sections of BPV-1-induced fibropapillomas by immunofluorescence. These MAbs were not reactive with intact BPV-1 particles or BPV-1 L1--galactosidase fusion proteins by ELISA or with -galactosidase by ELISA and Western blotting. The four MAbs detected viral structural proteins of M_r 76K, 68K and possibly 55K in purified BPV-1 preparations by Western blotting. Two of the four MAbs were cross-reactive with BPV-2-induced fibropapillomas. These findings suggest that (i) the BPV-1 L2 ORF encodes the minor capsid protein(s), (ii) the gene products of the BPV-1 L2 ORF have M_r values of 76K, 68K and possibly 55K, (iii) minor capsid epitopes are internal to the BPV-1 particle, and (iv) MAbs reactive with genetically engineered truncated BPV-1 L2 ORF gene products can distinguish between BPV-1 and BPV-2 productive infections.

Keywords: BPV-1, fusion proteins, L2 ORF

Received 24 October 1988; accepted 13 January 1989.

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