Abstract

Abstract Several adjuvant regimens were assessed for their ability to safely develop a high titer, high affinity antibody response in rabbits. Female, New Zealand White (NZW) rabbits were given five monthly 20 ug immunizations with a 25 KDa recombinant protein using one of four adjuvant programs: Adjulite® Complete and Adjulite® Incomplete Freund’s adjuvants,Alhydrogel® Aluminum hydroxide gel adjuvant in combination with oligodeoxynucleotides containing unmethylated CpG dinucleotides (Alum/CpG),Difco Freund’s adjuvant alternating with MPL+TDM+CWS orDifco Freund’s adjuvant alternating with Quil A supplemented with CpG-ODN. Sera samples taken following all booster immunizations were evaluated for antibody titer and relative affinity in a microtiter enzyme immunoassay by testing for antibody reactivity to limiting amounts of the administered antigen. Animals were also monitored during the immunization period for any reaction at the sites of injection. Results show that the Adjulite® animals elicited a higher average titer compared to animals immunized using any of the alternate strategies. Additionally, Adjulite® rabbits developed an average relative antibody affinity similar to that for the Alum/CpG animals and higher than either of the groups employing Difco Freund’s adjuvant. One Adjulite® rabbit developed a temporary nodule at the site of injection compared to none of the Alum/CpG animals. Two of the five animals using Difco Freund’s adjuvant developed a nodule, both of which progressed to lesions. This study demonstrates that Adjulite® adjuvant can serve as a safe alternative to other adjuvant regimens for the development of a high titer, high affinity antibody response in rabbits.

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