Hussein HA, Saber MS, Amer HM, Shalaby MA and Reda IM
Department of Virology, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.
A panel of monoclonal antibodies (Mabs) against the avian
reovirus reference strain S1133 was produced. Avian Reovirus (ARV) was
propagated on Vero cells, titrated and concentrated. The viral polypeptides
were identified using SDS-PAGE. Balb/C mice of 6-8 weeks old were immunized
using the concentrated reovirus antigen mixed with complete adjuvant. Booster
doses with the antigen mixed with and without adjuvant were then applied. The
mice spleenocytes were fused with Myeloma cells using polyethylene glycol.
Hybridoma cells that produce antibodies to ARV were selected using ELISA.
Positive hybridomas were cloned using limiting dilution to reveal subclones of
one-cell-origin. The produced Mabs were characterized using solid phase ELISA,
Immuno-binding, Immunofluorscent, Virus neutralization and Western blot
techniques. Three of the produced Mabs were specific to μB protein and one to
σB protein while the rest failed to recognize any of the ARV proteins in
Western blot. Non of them was neutralizing antibody.