The production and characterisation of dinitrocarbanilide antibodies raised using antigen mimics.
Polyclonal antibodies were produced to detect the coccidiostat nicarbazin. Due to structural constraints of the active component of nicarbazin, dinitrocarbanilide (DNC), three different compounds that shared a common substructure with DNC were used as antigen mimics. The compounds (N-succinyl-L-alanyl-L-alanyl-L-alanine 4-nitroanilide (SAN), L-glutamic acid gamma-(p-nitroanilide) (GAN) and p-nitrosuccinanilic acid (NSA)) were conjugated to a carrier protein and used in the immunisation of rabbits. Five different polyclonal sera were produced and consequently characterised. The antibodies exhibited an IC(50) range of 2.3-7.6 ng/ml using a competitive ELISA procedure. Serum from one rabbit, R555, exhibited an IC(50) of 2.9 ng/ml for DNC and cross-reactivity studies showed that this serum was specific for DNC and did not cross-react with other coccidiostats such as halofuginone, toltrazuril or ronidazole.
Veterinary Sciences Division, Department of Agriculture and Rural Development for Northern Ireland, Stoney Road, Northern Ireland, Belfast, UK. Lisa_Connolly@yahoo.com
Lisa
L
Connolly
Terence L
TL
Fodey
Steven R H
SR
Crooks
Philippe
P
Delahaut
Christopher T
CT
Elliott
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D000941:Antigens / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism; D002224:Carbanilides / Q000008:administration & dosage / Q000276:immunology* / Q000378:metabolism; D002352:Carrier Proteins / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D003049:Coccidiostats / Q000276:immunology; D003429:Cross Reactions; D004151:Dipeptides / Q000008:administration & dosage / Q000276:immunology; D005973:Glutamine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000276:immunology; D007106:Immune Sera / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007273:Injections, Intramuscular; D018716:Molecular Mimicry / Q000276:immunology; D009528:Nicarbazin / Q000276:immunology; D011817:Rabbits
D000941:Antigens; D002224:Carbanilides; D002352:Carrier Proteins; D003049:Coccidiostats; D004151:Dipeptides; D007106:Immune Sera; D005973:Glutamine; D009528:Nicarbazin; C053448:succinyl-alanyl-alanine-4-nitroanilide; C001300:gamma-glutamine-4-nitroanilide; C039493:4,4'-dinitrocarbanilide