title: Engineering of a broad specificity antibody for simultaneous detection of 13 sulfonamides at the maximum residue level.
abstract: Sulfa antibiotics (sulfonamides) are a group of molecules sharing the p-aminobenzenesulfonamide moiety. Sulfonamides are used in veterinary and human medicine. Sometimes, the meat or milk of medicated animals is contaminated with residual sulfonamides. Current analytical methods for sulfonamides are unfit for screening of food, because they are either too laborious, insensitive, or specific for a few sulfa compounds only. A rapid immunoassay for detection of all sulfas in a single reaction would thus be useful. Previously, we used protein engineering to improve the broad specificity of sulfa antibody 27G3. In this study, we improved the best mutant of the previous studies with site-directed mutagenesis. The new mutants recognized different sulfonamides with affinities sufficient for detection of all 13 tested sulfonamides below the MRL level. We furthermore demonstrated the functionality of one mutant in some real sample matrices.
authors.affiliation: Departments of Biotechnology and Bio-Organic Chemistry, University of Turku, Turku, Finland. teemu.korpimak@utu.fi
authors.forename: Teemu
authors.initials: T
authors.lastname: Korpimäki
authors.forename: Eeva-Christine
authors.initials: EC
authors.lastname: Brockmann
authors.forename: Outi
authors.initials: O
authors.lastname: Kuronen
authors.forename: Maija
authors.initials: M
authors.lastname: Saraste
authors.forename: Urpo
authors.initials: U
authors.lastname: Lamminmäki
authors.forename: Mika
authors.initials: M
authors.lastname: Tuomola
mesh_terms: D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity; D004350:Drug Residues / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis*; D015723:Gene Library; D007118:Immunoassay / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D013449:Sulfonamides / Q000032:analysis* / Q000276:immunology
chemical_list: D000906:Antibodies; D013449:Sulfonamides