Production of immunoglobulin Y against dog erythrocyte antigen (DEA) 1.1 from egg yolk

Abstract

The dog erythrocyte antigen (DEA) as a blood group antigen in dogs, especially DEA 1.1, has the highest
immunogenicity. IgY is an excellent antibody for use in immunological assays involving mammalian sera. The aim of
this study was to produce and purify IgY against DEA1.1 from chicken egg yolks for canine blood group detection.
Hens were immunized by intramuscular vaccination. IgY was achieved after three booster immunizations by DEA 1.1-
positive canine ghost cells. The extraction of total IgY from the egg yolk was carried out using precipitation by
polyethylene glycol 6000 and dialysis. The purified protein and yield of IgY were 102.52 ± 0.13 mg/ml and 25.13 ±
2.45%. Electrophoretic separation of egg yolk IgY showed other bands that appeared between the heavy and light
chains of IgY corresponding to molecular weights between 65 and 27 kDa. Canine red blood cells with DEA1.1 positive
which agglutinated with monoclonal antibody against DEA1.1 in a Rapid Vet H detection kit reacted with polyclonal
antibody against DEA1.1. Hemagglutination of three sample-polyclonal antibodies against DEA1.1 and two samples
of canine red blood cell with DEA1.1 positive was shown at titer 4. The purified IgY can be standardized and relatively
simple to perform, compared with standardization for blood typing. We successfully purified chicken IgY against
DEA1.1 from egg yolks of immunized hens, which can be used for diagnostic reagent for canine blood group
DEA1.1 detection. It is expected that IgY will play an increasing role in research and diagnosis in the future.