Qualitative analysis of antibody binding. An in vitro assay for the evaluation and development of vaccines.
We describe a rapid in vitro assay for the evaluation of in vivo properties of conjugate vaccines. Using human and murine monoclonal antibodies (mAb) specific for lipopolysaccharides (LPS), isolated from Pseudomonas aeruginosa, we determined in a competitive binding assay the amount of LPS or conjugate vaccine which was required to inhibit the antibody binding to LPS by 50% (I50 values). Furthermore, utilizing a murine burn wound sepsis model, we determined the potential of the same conjugates to induce protection in vivo against infection with the corresponding bacteria. Protective mAb have approximately 100-fold lower I50 values for preparations which are highly effective in inducing protection than for preparations which are ineffective. Furthermore, in the case of potent preparations it was noted that protective mAb exhibit similar I50 values for the conjugates and for the corresponding LPS. These results suggest that the fast and easily interpretable in vitro assay described may significantly facilitate the development and optimization of vaccines.
Bruderer
U
U
Swiss Serum and Vaccine Institute, Berne.
Fürer
E
E
Cryz
S J
SJ
Lang
A B
AB
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D001667:Binding, Competitive / Q000276:immunology*; D006801:Humans; D007114:Immunization; D000922:Immunotoxins / Q000008:administration & dosage / Q000276:immunology*; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D011552:Pseudomonas Infections / Q000517:prevention & control; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D011817:Rabbits
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D000922:Immunotoxins; D008070:Lipopolysaccharides; C029294:toxin A (Pseudomonas)