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. 2024 Dec 20;13(24):4137.
doi: 10.3390/foods13244137.

Development of a Colloidal Gold Immunochromatographic Assay Strip Using a Monoclonal Antibody for the Rapid Detection of Ofloxacin

Xiaolan Li  1 , Jin Huang  1 , Na Li  1 , Mahmoud Salah  2 , Shuoning Guan  1 , Wenwen Pan  1 , Ziyi Wang  1 , Xinghua Zhou  1 , Yun Wang  1
Affiliations

Development of a Colloidal Gold Immunochromatographic Assay Strip Using a Monoclonal Antibody for the Rapid Detection of Ofloxacin

Xiaolan Li et al. Foods. .

Abstract

The livestock industry uses ofloxacin, an antibiotic, to prevent several animal diseases; however, the overdose of ofloxacin used in animal farming treatments may appear in food products and cause some adverse human health effects. Hence, there is an immediate need to develop a method suitable for on site large-scale detection of ofloxacin residues in animal-derived foods. This study aimed to prepare a monoclonal antibody with high sensitivity and affinity for ofloxacin by re-synthesizing the ofloxacin hapten and synthesizing the corresponding complete antigen. The IC50 of the enzyme-linked immunosorbent assay (ic-ELISA) was 0.13 ng/mL, and the detection limit was 0.033 ng/mL. The visual detection limit of the established colloidal gold immunochromatographic test strip, for the visual detection of actual samples, was 1 ng/g. In summary, this work establishes a rapid detection method of ofloxacin residues on the basis of colloidal gold immunochromatography that is suitable for actual detection.

Keywords: immunochromatographic strip; monoclonal antibody; ofloxacin; rapid detection.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Chemical structure of OFL.
Figure 2
Figure 2
Modification of the hapten of OFL.
Figure 3
Figure 3
Preparation of immunogen and coating antigen ((a) OFL immunogen; (b) OFL coating antigen).
Figure 4
Figure 4
The basic principle of the colloidal gold immunochromatographic assay: (A) structure and (B) principle.
Figure 5
Figure 5
Mass spectrometry of the modified OFL.
Figure 6
Figure 6
The 1H NMR spectrum for the modified OFL.
Figure 7
Figure 7
UV–vis spectrum of OFL immunogen (a) and coating antigen (b).
Figure 8
Figure 8
Protein standard curve.
Figure 9
Figure 9
The screening results of hybridoma cells of OFL.
Figure 10
Figure 10
Subtype determination of mAb.
Figure 11
Figure 11
Affinity constant result of OFL antibody.
Figure 12
Figure 12
The optimization results of ELISA working conditions: blocking conditions (a), competition time (b), NaCl content (c), and pH (d).
Figure 13
Figure 13
The standard inhibition curves of OFL.
Figure 14
Figure 14
The characteristic images of colloidal gold particles: (a) UV–visible spectra, and (b) transmission electron microscopy images.
Figure 15
Figure 15
The images of colloidal gold immunochromatographic strip tests for OFL in (a) pork, (b) fish, and (c) chicken samples. (Note: the additional amounts of OFL standard in these samples are 1: 0 ng/g, 2: 1 ng/g, 3: 2 ng/g, 4: 4 ng/g).
See this image and copyright information in PMC

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