The accumulation of potato virus Y- (PVY) in tissue culture of Nicotiana tabacum (N. tabacum) was studied. Plants of N. tabacum variety, Samsun, inoculated with PVY-infected sap of potato variety, Cherie, were used for the virus accumulation. According to enzyme-linked immunosorbent assay (ELISA) results, virus showed the highest optical density (OD) on the 25^th day of inoculation. Murashige and Skoog medium containing kinetin 2 mg/l, 2.4-D 0.5 mg/l, indole-acetic acid 1 mg/l sucrose 2% agar 0.7%, in addition to the standard components, was used to induce callus culture from N. tabacum leave explants. ELISA results showed that the callus culture was able to maintain viral infection during four transplantations. Slightly and highly purified (Y-Cherie) virus preparations were obtained from the PVY-infected tissue culture. The slightly-purified antigens showed an OD approximately equal to the positive control in sandwich ELISA. The Y-Cherie antigen was detected as PVY necrotic strain. Specific to the virus polyclonal antibodies that reacted with a maximum 1/3200 titer of antigen in indirect ELISA were obtained in the result of the laboratory mouse immunisation.