Development of a monoclonal antibody-based cELISA for the analysis of sulfadimethoxine. 1. Development and characterization of monoclonal antibodies and molecular modeling studies of antibody recognition.
Sulfonamide antibiotics are used to treat a variety of bacterial and protozoan infections in cattle, swine, and poultry. Current residue methods for the analysis of sulfonamides in animal-based food products include bioassays, chromatographic methods (HPLC, GLC), and immunoassays. Most immunoassays have employed highly specific polyclonal antibodies. In this paper, we describe the isolation of monoclonal antibodies against sulfadimethoxine (SDM) that vary in their sensitivities and cross-reactivities against a large number of sulfonamides. The most sensitive monoclonal antibody, designated SDM-18, exhibits an IC(50) value for SDM of 1.53 ppb. Another monoclonal antibody, designated SDM-44, exhibits IC(50) values for six sulfonamides well below the established threshold level of 100 ppb for animal tissues. Molecular modeling studies of the cross-reactive drugs suggest that, depending on the monoclonal antibody, both steric and electronic features govern antibody binding. Due to the diversity of these monoclonal antibodies, it should be possible to design both compound- and class-specific monoclonal antibody-based immunoassays.
Muldoon
M T
MT
Food Animal Protection Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 2881 F&B Road, College Station, Texas 77845, USA.
Holtzapple
C K
CK
Deshpande
S S
SS
Beier
R C
RC
Stanker
L H
LH
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002417:Cattle; D002459:Cell Fusion; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D008956:Models, Chemical; D008958:Models, Molecular; D013412:Sulfadimethoxine / Q000032:analysis*
D000911:Antibodies, Monoclonal; D013412:Sulfadimethoxine