SELECTION OF AN ADJUVANT TO RAISE POLYCLONAL ANTIBODIES TO FOOT-AND-MOUTH DISEASE VIRUS IN RABBITS AND GUINEA-PIGS Alison Morris | Beatriz Sanz-Bernardo.

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Presentation transcript:

SELECTION OF AN ADJUVANT TO RAISE POLYCLONAL ANTIBODIES TO FOOT-AND-MOUTH DISEASE VIRUS IN RABBITS AND GUINEA-PIGS Alison Morris | Beatriz Sanz-Bernardo | Alison Burman | Anna Ludi The Pirbright Institute, Ash Road, Pirbright, Woking, GU24 0NF UK. Tel: +44 (0)1483 234455. Email: beatriz.sanz-bernardo@pirbright.ac.uk www.pirbright.ac.uk

Aims and Experimental Design Alternative to Freund’s adjuvant for the production of polyclonal antibodies. Good antibody titers but painful adverse effects. 3 adjuvant candidates (water-in-oil-emulsions): Adjuvant 1 (TitreMax-Gold). Adjuvant 2 (Montanide ISA50_V2). Adjuvant 3 (Stimune). Immunization protocol: Prime inoculation followed by a boost on day 28. Serum: day 0, day 22 and day 42 (end of experiment)  indirect ELISA.

Testing antisera to the homologous antigen Successful antibody production in all adjuvants. Montanide > Stimune > TitreMax-Gold. TitreMax-Gold (boost of soluble antigen without adjuvant). Further work to be done: Specificity ELISA.

Work in progress to refine the immunization protocol Adverse reactions To the prime injection: Significant skin inflammation in GP with TitreMax-Gold. To the boost: No reactions were observed in Group 1 – GP and rabbits - (boosted with antigen in PBS). An increase inflammation during the boost (compared to prime injection) in rabbits of Groups 2 and 3. In GP the adverse effects were better characterized by the pain response, which was greater in Groups 2 and 3. Adjuvant Species Number of animals Reason for Euthanasia TitreMax-Gold GP 2 Severe inflammation and pain of the injection site. Severe lesions upon necropsy. Rabbit Severe inflammation of the injection site. Severe lesions upon necropsy. Montanide ISA50 3 Moderate inflammation and pain on the injection site. Moderete lesions upon necropsy. Stimune 1 Pain on the injection site no associated with lesions produced by the inoculum. Work in progress to refine the immunization protocol