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The Thai Journal of Veterinary Medicine

Abstract

The dog erythrocyte antigen (DEA) as a blood group antigen in dogs, especially DEA 1.1, has the highest immunogenicity. IgYis 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 showedother bands that appeared between the heavy and light chains of IgY corresponding to molecular weights between 65 and 27kDa. 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 fromegg yolks of immunized hens, which can be used for diagnostic reagent forcanine blood group DEA1.1detection. It is expected that IgY will play an increasing role in research and diagnosis in the future

Publisher

Faculty of Veterinary Science, Chulalongkorn University

First Page

381

Last Page

387

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