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Isolation of porcine IgE, and preparation of polyclonal antisera
1Department of Veterinary Medicine, University of Bristol, Langford, UK.
Abstract:
High Titre, Ascaris specific, porcine reaginic antibody was induced by combining parasitic infestation and antigen challenge. The reagin was characterised as being heat labile at 56 degrees C, possessing long term skin sensitising activity, and as having physicochemical properties similar to IgE in other species. Porcine IgE was purified by conventional immunochemical techniques, and an epsilon chain isolate prepared using SDS-PAGE gel excision and electro elution. Antisera raised against the intact IgE molecule and its epsilon chain were rendered monospecific by immunoabsorption and characterised by reverse cutaneous anaphylaxis (RCA) testing, passive cutaneous anaphylaxis inhibition, immunohistology and Western blotting. These antisera are currently being used to study the regulation of IgE responses in the pig.