Goat serum containing highest antibody titers to FMD virus serotypes were processed separately via double precipitation and dialyzation. Fab components were purified and SDS PAGE was performed for confirmation. SDS-PAGE profiling of fragment antibody binding components of idiotype antibodies (IgG) after staining with Coomassie brilliant blue. L is for protein ladder while lane GA (group serotype Asia), GA1 (group serotype Asia1) and GO (group serotype O) are Fab components obtained from goat serum immunized against inactivated FMD virus serotypes. Band densities are indicating presence of more Fab components in the group injected with FMD virus serotype Asia 1 while least in group representing serotype O. 

(A) Detection of anti-idiotype antibodies: Purified idiotype Fab components from respective groups were adjuvanted in Montanide and injected in layer birds. Eggs were processed for purification of anti-idiotype antibodies to Fab components of polyclonal idiotype antibodies against three FMD virus serotypes. Presence of anti-idiotype antibodies were confirmed via agar gel immune precipitation test. A clear Line of precipitation in the figure between idiotype antigen placed in the central well and egg yolk containing anti-idiotype antibody on the 0.9% agarose gel plate after incubation of 48 hrs from the group II received the idiotype antigen containing protein concentration of 10mg/mL. (B). Confirmation of anti-idiotype antibodies: The anti-idiotype proteins mimicking FMD virus were disrupted on SDS-PAGE and transferred onto the nitrocellulose membrane. The anti-idiotype proteins mimicking FMD virus were detected at 27KDa using VP1 antibody to FMD virus and GAPDH was used as loading control.
 

Rabbits were divided in four groups as RO (Group of rabbits for serotype O), RA (Group of rabbits for serotype A), RA1(Group of rabbits for serotype Asia 1) and RC (Group of rabbits for control). Anti-idiotype bodies purified from egg yolks were adjuvanted in Montanide and injected in rabbits. Viral % inhibitory were measured via C-ELISA and results are expressed indicating time dependent inhibitory levels.