首页|Development of Fc-specific multi-biotinylated antibodies via photoreactive tandem AviTag repeats for the ultrasensitive determination of ochratoxin A
Development of Fc-specific multi-biotinylated antibodies via photoreactive tandem AviTag repeats for the ultrasensitive determination of ochratoxin A
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NSTL
Elsevier
Ochratoxin A (OTA) is a common food contaminant that seriously threatens the safety and health of consumers. Here, an indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (icBA-ELISA) was developed for the sensitive detection of OTA by employing Fc-specific multi-biotinylated antibodies. An Fcbinding protein (Z-domain) carrying a photo-cross-linker (p-benzoylphenylalanine, Bpa) fused with tandem AviTag repeats, viz. Z(Bpa)-(AviTag)(n), was first constructed using the aminoacyl-tRNA synthetase/suppressor tRNA technique. Biotin molecules were covalently conjugated to antibodies mediated by the photoreactive biotinylated Z(Bpa)-(AviTag)(n), resulting in the Fc-specific multi-biotinylated antibodies (IgG-*Z-B-n conjugates) that were used to establish icBA-ELISA. Among the IgG-*Z-B-n-based icBA-ELISA, the IgG-*Z-B-5 conjugates exhibited a half maximal inhibitory concentration of 0.0519 ng mL(-1) and a limit of detection of 0.00944 ng mL(-1) for OTA. The sensitivity was improved 4.8 and 4.7 times compared with that of the chemical-coupled IgG-biotin conjugates. Thus, the proposed icBA-ELISA for the sensitive detection of OTA has remarkable potential applications for the detection of various toxins in food.
NSTL
Elsevier
