Induction and rapid screening of monoclonal antibodies against cyclosporin A.
Cyclosporin C-hemisuccinate was coupled to thyroglobulin by the mixed anhydride method. Cyclosporin C was used instead of cyclosporin A (CsA) because of lack of functional groups of CsA. The resulting protein-cyclosporin C conjugate allowed us to induce high antibody titers also against cyclosporin A in rabbit and mice. Polyclonal and monoclonal antibodies were prepared following standard procedure. Since no standard methods for screening and quantification of anti-CsA-antibodies were available, two methods were adapted: (a) liquid phase radio assay (RA) and (b) solid phase enzyme-linked immunoassay (ELI-SA). For the former procedure inactivated charcoal was applied to separate the antibody-bound and the unbound CsA. CsA-coated PVC microtiter plates were used for the latter.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D003524:Cyclosporins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D007163:Immunosorbent Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D003524:Cyclosporins