Production and Selection of a Monoclonal Antibody specific to a Japanese Necrotic Strain of Potato Virus Y

摘要

A Japanese necrotic strain of Potato virus Y (PVY-T) is serologically related with an ordinary strain of PVY (PVY-O); therefore, a rabbit antiserum against PVY-T that was thoroughly absorbed with the PVY-O antigen has been used for detection of PVY-T.To detect PVY-T more precisely from tobacco leaves or potato young buds by serological methods, I produced monoclonal antibodies specific to PVY-T. A panel of monoclonal antibodies was raised against PVY-T virion using a standard procedure. The bindingproperties of individual antibodies to PVY-T and PVY-O were analyzed by indirect enzyme-linked immunosorbent assay. Twenty-eight out of 238 hybridoma cell lines obtained in the experiment produced antibodies specific to PVY-T. The 5 T 2 cell line that showed the highest titer among these specific lines was injected back into the mouse peritoneum and ascitic fluid was obtained. The 5 T 2 antibody belongs to the IgG 2 a subclass and has a kappa light chain. The nucleotide sequence of the coat protein geneof PVY-T was determined and the 5 T 2 antibody was suggested to react to the N-terminal region of the coat protein.