DEVELOPMENT OF MONOCLONAL-ANTIBODIES FOR MEASUREMENT OF IMMUNOGLOBULIN-G ANTIBODY-RESPONSES IN BLUE AND GOLD MACAWS (ARA-ARARAUNA)

Objectives-To produce monoclonal antibodies (MAB) with specificity for the heavy chain of macaw IgG; to incorporate these MAB into an ELISA to measure IgG responses of macaws inoculated with bovine serum albumi n (BSA); and to evaluate the antigenicity of BSA in Blue and Gold Maca ws. Animals-Four 1-year-old Blue and Gold Macaws, 2 males and 2 female s. Procedure-1 male and 1 female 1 were randomly assigned to each of 2 study groups. Group-1 birds were inoculated with 200 mu g of BSA on d ays 0, 14, 28, and 42. Group-2 birds were inoculated with 200 mu g of BSA on days 0 and 28. Blood was collected weekly for measurement of an ti-BSA titer. Hybridomas were prepared from mice immunized with Scarle t Macaw (Ara macao) IgG purified by salt precipitation and gel chromat ography. Specificity for IgG of Scarlet Macaw and other macaw species was confirmed by ELISA and western blot analysis, Hybridoma HL544 was cloned and the antibody purified. Following biotinylation, MAB HL544 w as incorporated into an ELISA that measured IgG responses of macaws in oculated with BSA. Results-Adult Blue and Gold Macaws developed strong primary and secondary anti-BSA antibody titers 14 days after inoculat ion with 200 mu g of BSA. An inoculation interval of 28 days resulted in stronger secondary responses than an interval of only 14 days. Conc lusions-MAB specific for macaw immunoglobulins can be used in ELISA to evaluate the humoral immune responses of macaws. 1-year-old Blue and Gold Macaws developed strong anti-BSA titer when inoculated with 200 m u g of BSA. An inoculation interval of 28 days resulted in stronger se condary responses than did an interval of only 14 days. Clinical Relev ance-These MAB, the first reported to have specificity for a psittacin e antibody class, will je useful in the evaluation of psittacine antib ody responses and in the development of psittacine diagnostic assays.