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. 2023 May 16:11:1196043.
doi: 10.3389/fbioe.2023.1196043. eCollection 2023.

Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies

Yongming Huang  1 , Aidi Xu  1 , Yang Xu  1 , Huijuan Wu  1 , Menghan Sun  1 , Lakshani Madushika  1 , Rongzhi Wang  1 , Jun Yuan  1 , Shihua Wang  1 , Sumei Ling  1
Affiliations

Sensitive and rapid detection of tetrodotoxin based on gold nanoflower-and latex microsphere-labeled monoclonal antibodies

Yongming Huang et al. Front Bioeng Biotechnol. .

Abstract

Tetrodotoxin (TTX) could result in serious diseases due to its extremely high neurotoxicity. Thus, it is of great importance to measure TTX for food safety. In this study, an anti-TTX monoclonal antibody with good specificity and high affinity was used to develop the immunochromatographic test strips (ICTS). Gold nanoflower (AuNF) with multiple branches and latex microsphere (LM) with large particle size as signal reporters were employed for improving the sensitivity of test strips. Both AuNF and LM probes are stable, and the developed ICTS were specific to TTX, demonstrating no cross-reactivity with other marine toxins. The linear range of AuNF- and LM-based strips for TTX was 9.49-330.98 ng/mL and 5.40-443.19 ng/mL, respectively. The limit of detection (LOD) of AuNF- and LM-based strips was determined to be 9.49 ng/mL and 5.40 ng/mL, respectively. In summary, the developed ICTS based on AuNF and LM signal probes displayed enhancement of sensitivity and provided rapid and specific detection of TTX.

Keywords: detection; gold nanoflower; immunochromatography strip; latex microsphere; monoclonal antibody; tetrodotoxin.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Synthesis and characterization of AuNF. (A) Synthesis process of AuNFs and AuNF probes. (B) Transmission electron microscopy (TEM) result of AuNFs. (C) Ultraviolet-visible absorption spectra of AuNFs (red) and AuNF probes (blue).
FIGURE 2
FIGURE 2
Representation of the AuNF-based strip (n = 3). (A) Schematic diagram of the developed AuNF-based strip. (B) Specificity of the AuNF-based strip by adding different marine toxins. (C) TTX with different concentrations effects on the color development of C and T lines on AuNF-based strips. (D) Immunochromato-reader absorbance values associated with the results of (C) for AuNF-based strips. (E) Linear range of TTX detection by AuNF- based strips.
FIGURE 3
FIGURE 3
Synthesis and characterization of LM. (A) Synthesis process of LM probes. (B) TEM result of LMs.
FIGURE 4
FIGURE 4
Representation of the LM-based strip (n = 3). (A) Specificity of the LM-based strip. (B) TTX with different concentrations was added to LM-based strips to test the sensitivity results. (C) T value of LM-based strips using the immunochromato-reader corresponding to the results of (B). (D) Linear range of TTX detection by LM-based strips.
FIGURE 5
FIGURE 5
Stability of AuNF/LM-based strips (n = 3). (A) Stability results of AuNF-based strips. (B) Stability results of LM-based strips. (C) T + C values of the developed AuNF-based strip. (D) T + C values of the LM-based strip.
FIGURE 6
FIGURE 6
Real sample testing of the established AuNF/LM-based strips (n = 3). (A) Detection results of real samples determined by AuNF-based strips. (B) Testing results of real samples determined by LM-based strips.
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