Potential of the sulfobetaine detergent Zwittergent 3-12 as a desorbing agent in biospecific and bioselective affinity chromatography.

The isolation in our laboratories of several antigens of interest from sporulated oocysts of Eimeria species by bioselective adsorption on matrices containing immobilized antigen-specific immunoglobulins IgG was initially unsuccessful. The preparations serving as source materials for these antigens contained low levels of the zwitterionic sulfobetaine detergent, Zwittergent 3-12. Since usually immunoaffinity processes are carried out in the presence of various detergents, we were surprised, subsequently, to find this detergent to be the cause of the problem in that it prevented antigen-antibody binding. These findings led us to study the potential role of Zwittergent 3-12 as an eluting agent from matrices holding bioselectively adsorbed materials. The results of seven case studies are presented in this paper and include experiments with beta-D-galactosidase adsorbed biospecifically and bioselectively on matrices via either specific antibody or inhibitor analogue. In all cases, Zwittergent 3-12 proved to be an effective desorbing agent.

Passive protection of chickens against Eimeria tenella infection by monoclonal antibody.

Monoclonal antibodies reactive with the surface of Eimeria tenella sporozoites were produced in mice. This paper concerns one of these monoclonal antibodies, designated 1073.10, which agglutinated sporozoites in vitro and lysed the parasite in the presence of complement. This treatment neutralized sporozoite infections when the treated parasites were injected into the ceca of normal chickens. Passive transfer of ammonium sulfate-precipitated 1073.10 ascites fluid into 2- to 3-day-old or 3-week-old chickens conferred protection against challenge infection with E. tenella. These studies show that serum antibody may play a role in immunity to coccidiosis and that the sporozoite surface epitope recognized by 1073.10 is a possible vaccine candidate antigen.

Eimeria tenella: quantitative in vitro and in vivo studies on the effects of mouse polyclonal and monoclonal antibodies on sporozoites.

Murine, polyclonal and monoclonal antibodies, raised against sporozoites of Eimeria tenella, were tested for their ability to neutralize sporozoite infectivity in vitro and in vivo. Neutralization was effected via three mechanisms. Firstly, sporozoites fixed complement, at low titres, and lysis occurred by the alternative pathway of complement activation. Secondly, in the absence of complement activity, the murine heat-inactivated, hyperimmune antiserum neutralized sporozoites at relatively low titres. At high titres, even though sporozoites were agglutinated, neither the heat-inactivated hyperimmune antiserum nor the monoclonal antibody neutralized sporozoites. Finally, in the presence of complement and specific antibodies, at titres which by themselves would not neutralize sporozoites, neutralization was effected due to lysis via the classical pathway of complement activation.

Eimeria tenella (Eucoccidiorida): a quantitative assay for sporozoite infectivity in vivo.

A quantitative technique for the assessment of sporozoite infectivity in vivo, using intra-cecal inoculation of Eimeria tenella sporozoites, has been developed. Evaluation of the infection using cecal lesion scores and oocyst counts showed that this technique should be useful for the quantitation of sporozoite viability and thus for the anti-sporozoite activity of different treatments prior to inoculation. Pre-treatment of sporozoites with heat-inactivated hyperimmune antisera neutralized sporozoite infectivity in vivo and indicated that antibodies in the absence of complement inhibited sporozoite infectivity in vivo.