A strategy for inducing an immune response against Androctonus australis scorpion venom toxin I in mice. Production of high-affinity monoclonal antibodies and their use in a sensitive two-site immunometric assay.
Scorpion neurotoxins acting on ion channels share some structural features but differ in antigenic and immunogenic properties. They are highly structured peptides, 60-70 amino acids long. Monoclonal antibodies have been obtained for Androctonus australis hector scorpion venom neurotoxin II (AahII) and a nontoxic synthetic analog ((Abu)(8) AahII). In this study, no antibody response was elicited in mice of various strains injected with AahI, the other important toxin of the venom, in a native or an inactive ((Abu)(8) AahI) form. We found that AahI was only immunogenic in BALB/c or C57BL/6 mice if it was coupled to a carrier protein. The helper protein molecule could be BSA, KLH, or the nontoxic analog of AahII. We obtained a panel of high-affinity mAbs with these immunogens. Two of these mAbs, including the very high-affinity antibody 9C2 (K(D)=0.11x10(-11) M), were used to set up a two-site ELISA, sensitive enough for the quantification of AahI in the biological fluids of envenomed animals. The detection limit of the assay was 75 pg/ml.
Devaux
Christiane
C
Laboratoire de Biochimie-Ingénierie des Protéines, CNRS UMR 6560, Faculté de Médecine-Nord, Bd P. Dramard, 13916 Marseille Cedex 20, France. devaux.c@jean-roche.univ-mrs.fr
Clot-Faybesse
Olivier
O
Pugnière
Martine
M
Mani
Jean Claude
JC
Rochat
Hervé
H
Granier
Claude
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000997:Antivenins / Q000276:immunology; D002352:Carrier Proteins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009498:Neurotoxins / Q000276:immunology*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D012604:Scorpion Venoms / Q000276:immunology*; D012605:Scorpions / Q000276:immunology*; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D000997:Antivenins; D002352:Carrier Proteins; D007074:Immunoglobulin G; D009498:Neurotoxins; D012604:Scorpion Venoms; C065314:scorpion toxin I, Androctonus