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Anal Biochem ; 409(1): 14-21, 2011 Feb 01.
Article in English | MEDLINE | ID: mdl-20920457

ABSTRACT

Recombinant sarafloxacin-recognizing antibody was engineered with the use of novel fluoroquinolone (FQ) derivatives. A monoclonal FQ antibody, 6H7, was targeted to random mutagenesis to broaden the specificity of the antibody in development of a generic assay for FQ antibiotics. Engineering involved the synthesis of different small-sized FQ molecules to immobilize and detect the mutant antibodies. Selections with labeled FQs resulted in several mutant antibodies with increased affinity or wider specificity toward different FQs. The best characterized mutant antibody was capable of recognizing seven of eight targeted FQs below maximum residue limits set by the European Union. The results are promising in regard to the development of a multiresidue immunoassay for FQs based on a single antibody.


Subject(s)
Anti-Bacterial Agents/analysis , Antibodies, Monoclonal/immunology , Fluoroquinolones/analysis , Immunoassay/methods , Protein Engineering , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/immunology , Antibodies, Monoclonal/genetics , Antibodies, Monoclonal/metabolism , Antibody Specificity , Biotinylation , Fluoroquinolones/chemical synthesis , Fluoroquinolones/immunology , Mutagenesis , Peptide Library , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Structure-Activity Relationship
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