Comparison of two immunization routes for the production of chicken antibodies against fructose-1,6-bisphosphatase

The present study compares the intramuscular (IM) and subcutaneous (SC) inoculation routes for antibody production in hens. Six laying Leghorn hens were separated in two identical inoculation groups and immunized with purified Fructose-1,6-bisphosphatase (FBPase) from pig kidney extract. The hens were inoculated at two and four week intervals with an emulsified mixture of purified FBPase and Freund's complete adjuvant (first inoculation) and Freund's incomplete adjuvant (second and third inoculations). The egg yolk antibodies were purified and titrated with immune dot blot. High amounts of antibody were produced by both hen groups and the titers were comparable. However, in each inoculation group we found some differences which we attributed to individual variations. These results show that both routes of inoculation (SC and IM) are equally effective in generating hen antibodies against a cytosolic enzyme. Moreover, the immune-blot analysis revealed that the antibodies anti-FBPase obtained from intramuscular and subcutaneous inoculations recognized the enzyme in rat liver extract containing much more than 50 different proteins. These results demonstrated that the antibodies produced by these two routes of inoculation are highly specific.