Production of highly specific monoclonal antibodies to monensin and development of a microELISA to detect this antibiotic.
Monensin, a polyether antibiotic of molecular weight 671 Da, was converted into a hemisuccinate and covalently linked to bovine serum albumin via the mixed anhydride method. Using this immunogen, polyclonal anti-monensin antibodies were raised in rabbits and monoclonal antibodies were prepared from mice. The specificity of the anti-monensin antibodies was examined by using several structural analogues as the immunogen and by performing direct binding and competitive microELISA assays on Terasaki plates. Rabbit polyclonal antibodies had a dissociation constant (KD) of 5.5 x 10(-8) M for monensin and reacted with nigericin, an antibiotic structurally related to monensin. In contrast, a mouse monoclonal antibody, 2H8, reacted only with monensin and had a much lower KD = 3 x 10(-8) M for monensin. Monoclonal antibody 2H8 was used to develop a competitive microELISA able to detect as little as 5 ng/ml of monensin in solution which corresponds to 75 pg or 110 fmol of this hapten per Terasaki well.
CNRS URA 359, Département d'Immunologie, Institut Pasteur, Paris, France.
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Pauillac
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Halmos
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Labrousse
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Antonakis
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Avrameas
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D006241:Haptens; D007118:Immunoassay; D051379:Mice; D008985:Monensin / Q000032:analysis* / Q000276:immunology*; D011817:Rabbits
D000911:Antibodies, Monoclonal; D006241:Haptens; D008985:Monensin