Generation of class-selective monoclonal antibodies against the penicillin group

被引:28
作者
Cliquet, P
Cox, E
Van Dorpe, C
Schacht, E
Goddeeris, BM
机构
[1] State Univ Ghent, Fac Vet Med, Lab Vet Immunol, B-9820 Merelbeke, Belgium
[2] Katholieke Univ Leuven, Fac Agr & Appl Biol Sci, Lab Physiol & Immunol Domest Anim, B-3001 Heverlee, Belgium
[3] State Univ Ghent, Fac Sci, Organ Chem Lab, B-9000 Ghent, Belgium
关键词
beta-lactam antibiotics; immunogenicity; enzyme-linked immunosorbent assay; monoclonal antibodies;
D O I
10.1021/jf001428k
中图分类号
S [农业科学];
学科分类号
09 ;
摘要
To develop a penicillin-specific ELISA, different attempts were made to obtain monoclonal antibodies specific for the common structure of penicillins. Ampicillin was coupled to different carrier proteins (bovine serum albumin, chicken ovalbumin, and thyroglobulin) to render it immunogenic. Different coupling methods were compared: two methods using a cross-linker (glutaraldehyde or a succinimide ester), one carbodiimide-mediated coupling method, and one method without any cross-linker or mediator molecule (physiological binding). Mice were immunized with the conjugates intraperitoneally or in the footpad. A screening ELISA was developed to detect anti-ampicillin antibodies in sera. Specificity and affinity of the antibodies were demonstrated by inhibiting their binding with a 10 mM solution of ampicillin. No difference could be observed using electrofusion or PEG-mediated fusion. For the production of the monoclonals, an intraveneous final boost gave antibodies with better specificity and affinity than an intraperitoneal final booster injection. At least one anti-ampicillin monoclonal antibody (19C9) cross-reacts with penicillin G, oxacillin, dicloxacillin, and carbenicillin, and not with sulfanilamide, chloramphenicol, neomycin, and streptomycin, and is therefore considered interesting for developing a penicillin-specific ELISA.
引用
收藏
页码:3349 / 3355
页数:7
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