Production of a rabbit anti-cockatiel immunoglobulin G and characterization of its cross-reactivities with immunoglobulin G of other psittacine species

The purpose of this work was to produce rabbit anti-cockatiel immunoglobuli n G (IgG) and compare its cross-reactivity with sera from eight other psitt acine birds: Quaker parakeet, budgerigar, green-wing macaw, blue-fronted Am azon parrot, eclectus parrot, African grey parrot, Patagonian conure, Moluc can cockatoo. Cockatiel IgG did not bind to protein A or G; therefore, thes e proteins could not be used in column chromatography to isolate the IgG. A combination of serum IgG precipitation by ammonium sulfate and yolk IgG ex traction from egg was loaded in sodium dodecyl sulfare-polyacrylamide gel u pon which the IgG was resolved by electrophoresis. The resolved Ige in sodi um dodecyl sulfate-polyacrylamide gel was stained with Coomassie blue, cut, crushed in phosphate-buffered saline, and injected into rabbits. The rabbi t anti-cockatiel IgG produced in this way reacted with a single protein in gel immunodiffusion assay with all nine psittacine bird sera but not with t hose of chicken and ostrich. Immunoelectrophoresis confirmed the cross-reac tivity of different psittacine sera with the anci-cockatiel IgG serum but n ot with ostrich and chicken sera. This antiserum detected antibody response s in sera from cockatiels vaccinated against chlamydial major outer membran e protein in an immunoblot assay.