Production, characterization, and cross-reactivity studies of monoclonal antibodies against the coccidiostat nicarbazin.
A cELISA was developed for the coccidiostat nicarbazin. On the basis of previous computer-assisted molecular modeling studies, p-nitrosuccinanilic acid (PNA-S) was selected as a hapten to produce antibodies to 4,4'-dinitrocarbanilide (DNC), the active component of the coccidiostat nicarbazin. Synthesis is described for the hapten [p-nitro-cis-1,2-cyclohexanedicarboxanilic acid (PNA-C)] used in a BSA conjugate as a plate coating antigen. Monoclonal antibodies (Mabs) were isolated that compete with nicarbazin, having IgM(kappa) isotype. Because of the lack of water solubility of nicarbazin, N,N-dimethylformamide (DMF) (3%, v/v) and acetonitrile (ACN) (10%, v/v) were added to the assay buffer to achieve solubility of nicarbazin and related compounds. The Nic 6 Mabs had an IC(35) value for nicarbazin of 0.92 nmol/mL, with a limit of detection of 0.33 nmol/mL. Nic 6 exhibited high cross-reactivity for PNA-S and PNA-C, and 3-nitrophenol, 4-nitrophenol, and 1-(4-chlorophenyl)-3-(4-nitrophenyl) urea. However, Nic 6 had little or no cross-reactivity with 15 other related compounds.
Beier
R C
RC
Southern Plains Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 2881 F & B Road, College Station, Texas 77845-4988, USA. RCBeier@ffsru.tamu.edu
Ripley
L H
LH
Young
C R
CR
Kaiser
C M
CM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D002224:Carbanilides / Q000276:immunology; D002460:Cell Line; D003049:Coccidiostats / Q000276:immunology*; D003198:Computer Simulation; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D009528:Nicarbazin / Q000737:chemistry / Q000276:immunology*; D012995:Solubility
D000911:Antibodies, Monoclonal; D002224:Carbanilides; D003049:Coccidiostats; D006241:Haptens; D009528:Nicarbazin; C039493:4,4'-dinitrocarbanilide