A novel optical immunosensor for the screening of ampicillin (Amp) residues has been developed. The biosensor is based on fiber optic particle plasmon resonance detection and uses an enhancement method called as fiber optic nanogold-linked immunosorbent assay (FONLISA) for the sensitive detection of antibiotics. A commercial antibody which had a higher affinity for ampicillin than for other β-lactam antibiotics was chosen. A surface competitive binding assay was used in which a fixed concentration of antibiotic-conjugated gold nanoparticles (AuNPs) competes with free unlabeled antibiotic molecules to measure the amount of binding with antibody molecules immobilized on an optical fiber. The synthesis of the 11-mercaptoundecanoic acid (MUA)-ampicillin conjugate facilitates the attachment of the Amp molecules to AuNPs via MUA which acts as a linker between them. This AuNP–Amp conjugate was then used for the detection of β-lactam antibiotics. The practical limit of detection obtained for Amp was 0.74 ppb (7.4 × 10−10 g/mL) which is lower than the recommended maximum residue limit (MRL) for β-lactams. The method also shows a wide linear range of 4 orders. Its applicability to the determination of ampicillin in spiked milk samples has been demonstrated with good recovery and reproducibility.
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We thank Prof. Bor-Cherng Hong for helpful advice on the synthesis and characterization of Amp–MUA conjugate.
This work was supported by the Ministry of Science and Technology of Taiwan (Grant no. MOST 105-2113-M-194-009-MY3) and Center for Nano Bio-Detection from the Featured Research Areas College Development Plan of National Chung Cheng University.
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Electronic supplementary material
Sensorgram of nonspecific adsorption test by direct assay, sensorgram and calibration graph by direct assay, TEM image and UV-vis spectra of the prepared AuNPs, FTIR spectra of Amp and the synthesized Amp-MUA conjugate, calibration graph for quantitation of IgG on fiber surface, and sensorgram for ampicillin detection at concentration near practical LOQ by FONLISA are provided as supplementary information.
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Chaudhari, P.P., Chau, L., Tseng, Y. et al. A fiber optic nanoplasmonic biosensor for the sensitive detection of ampicillin and its analogs. Microchim Acta 187, 396 (2020). https://doi.org/10.1007/s00604-020-04381-w
- Fiber optic
- Gold nanoparticle