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Title: Generation of group-specific antibodies against sulfonamides
Authors: Cliquet, P ×
Cox, E
Haasnoot, W
Schacht, E
Goddeeris, Bruno #
Issue Date: 2003
Publisher: AMER CHEMICAL SOC
Series Title: Journal of Agricultural and Food Chemistry vol:51 issue:20 pages:5835-5842
Conference: date:State Univ Ghent, Fac Vet Med, Lab Vet Immunol, B-9820 Merelbeke, Belgium; Katholieke Univ Leuven, Fac Agr & Appl Biol Sci, Lab Physiol & Immunol Domest Anim, B-3001 Heverlee, Belgium; RIKILT, State Inst Qual Control Agr Prod, NL-6708 PD Wageningen, Netherlands; State Univ Ghent, Fac Sci, Chim Organ Lab, B-9000 Ghent, Belgium
Abstract: To develop a sulfonamide-specific ELISA, different attempts were made to obtain monoclonal antibodies specific for the common structure of sulfonamides. In a first approach, sulfanilamide was linked to albumins using glutaraldehyde or a succinimide ester as cross-linker. A weak immune response or none at all was induced after immunization of mice with those conjugates. High antibody titers were obtained with conjugates where sulfanilamide was linked to albumins or casein (azocasein) with a diazotation reaction. However, the antibodies were only highly specific for the bound sulfanilamide molecule. In a second approach, sulfonamide-protein conjugates were used in which the sulfonamide molecule is linked at its side chain, leaving the common structure of sulfonamides unchanged. Three sulfonamide derivatives (S, TS, and PS, previously described in the literature) containing a carboxyl group in their side chain were linked to proteins using a carbodiimide mediated reaction. Immunization with the S-conjugates led to high antibody titers, but the antibodies were only highly specific for the bound S-molecule. Group-specific antibodies were obtained after immunization with the PS- and TS-conjugates. It was described that immunization with PS-conjugates lead to the recognition of other sulfonamides (sulfamethazine, -merazine, -diazine, and -dimethoxine) that are not well recognized by antibodies induced after immunization with TS-conjugates. Therefore, we tried to guide the immune response in the direction of recognition of the common structure of sulfonamides by immunizing the animals alternately with PS- and TS-conjugates. The polyclonal antibodies of the mice indeed had a broader specificity, but the specificity of the monoclonals obtained after fusion experiments was not influenced. Immunization with TS-conjugates seemed sufficient to obtain sulfonamide-specific monoclonal antibodies. With the best monoclonal (mAb 3B5B10E3) two competitive inhibition (ci) ELISA's were developed: one coated with antigen and the other coated with the monoclonal antibody. Sulfadiazine, -dimethoxine, -thiazole, -pyridine, and -methoxazole were detected in both ELISA's at their MRL-value (100 ppb) in buffer solution. Sulfadiazine, sulfathiazole, and sulfamethoxazole, could even be detected at 10 ppb.
ISSN: 0021-8561
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Division of Gene Technology (-)
× corresponding author
# (joint) last author

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