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. 2024 Jul 15;14(7):342.
doi: 10.3390/bios14070342.

Magnetic Immunoassay Based on Au Pt Bimetallic Nanoparticles/Carbon Nanotube Hybrids for Sensitive Detection of Tetracycline Antibiotics

Jianxia Lv  1 , Rui Huang  2   3 , Kun Zeng  2   3 , Zhen Zhang  2   3
Affiliations

Magnetic Immunoassay Based on Au Pt Bimetallic Nanoparticles/Carbon Nanotube Hybrids for Sensitive Detection of Tetracycline Antibiotics

Jianxia Lv et al. Biosensors (Basel). .

Abstract

Misusage of tetracycline (TC) antibiotics residue in animal food has posed a significant threat to human health. Therefore, there is an urgent need to develop highly sensitive and robust assays for detecting TC. In the current study, gold and platinum nanoparticles were deposited on carbon nanotubes (CNTs) through the superposition method (Au@Pt/CNTs-s) and one-pot method (Au@Pt/CNTs-o). Au@Pt/CNTs-s displayed higher enzyme-like activity than Au@Pt/CNTs-o, which were utilized for the development of sensitive magnetic immunoassays. Under the optimized conditions, the limits of detection (LODs) of magnetic immunoassays assisted by Au@Pt/CNTs-s and Au@Pt/CNTs-o against TCs could reach 0.74 ng/mL and 1.74 ng/m, respectively, which were improved 6-fold and 2.5-fold in comparison with conventional magnetic immunoassay. In addition, the measurement of TC-family antibiotics was implemented by this assay, and ascribed to the antibody used that could recognize TC, oxytetracycline, chlortetracycline, and doxycycline with high cross-reactivity. Furthermore, the method showed good accuracy (recoveries, 92.1-114.5% for milk; 88.6-92.4% for pork samples), which also were applied for determination of the targets in real samples. This study provides novel insights into the rapid detection of targets based on high-performance nanocatalysts.

Keywords: immunoassay; nanozyme; rapid detection; tetracycline.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The diagram of magnetic immunoassay assisted by Au@Pt/CNTs.
Figure 2
Figure 2
Characterizations of Au@Pt/CNTs nanohybrid. (a–c) SEM, TEM, and EDS images of Au@Pt/CNTs-s; (d–f) SEM, TEM, and EDS images of Au@Pt/CNTs-o.
Figure 3
Figure 3
(a) Identification of enzyme−like probes: 1 uncarboxylated MWCNTs; 2 Au@Pt; 3 carboxylated MWCNTs; 4 Au@Pt/CNTs-s; 5 Au@Pt/CNTs-o. (b) Kinetic curve of enzymatic reaction; the double reciprocal plot of the activity of Au@Pt/CNTs-s (c,d) and Au@Pt/CNTs-o (e,f).
Figure 4
Figure 4
Optimization of reaction parameters: (a) concentration of GAM; (b) volume of IMB-GAM; (c) blocking buffer. TEM images of combination between IMB and Au@Pt/CNTs-s (d) or Au@Pt/CNTs-o (e); IMBs are circled in red. Optimization of the volume of probes (f).
Figure 5
Figure 5
Optimal conditions in magnetic immunoassay assisted by Au@Pt/CNTs-s (a–d) and Au@Pt/CNTs-o (e–h).
Figure 6
Figure 6
Standard curves of magnetic immunoassays assisted by HRP-TC, Au@Pt/CNTs-s, and Au@Pt/CNTs-o.
Figure 7
Figure 7
Cross-reactivity of magnetic immunoassays assisted by Au@Pt/CNTs-s.
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