Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Detecting Tetrodotoxin

Qing Ping Zhong; Yan Ting Liu; Yuan Ming Sun; Hoi Fu Yu; Hong Tao Lei

doi:10.4028/www.scientific.net/AMR.466-467.220

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HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 466-467Development of Monoclonal Antibody-Based...

Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Detecting Tetrodotoxin

Abstract:

This study aimed to develop a method for the detection of tetrodotoxin (TTX) based on monoclonal antibody (McAb). The hapten TTX was linked to carrier protein keyhole limpet hemocyanin (KLH) as immunogen, and linked to ovalbumin (OVA) as coating antigen by the Mannich method. Then the 6~8 weeks Babl/c mice were immunized. After cell amalgamation, a cell line with high specificity and sensitivity was obtained, and McAb was produced. The indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed for detecting TTX. The optimized working conditions of the icELISA were 4.0 μg/mL coating antigen, 0.75 μg/mL McAb, 45 min competitive time, at room temperature (20~25 °C). The IC₅₀ value of this method was 24.0 ng/mL, the working ranges were 5.2~107.6 ng/mL, the intra-assay and inter-assay coefficient of variation (CV %) were 4.2 and 4.5, respectively. This investigation will benefit the assay kit development for detecting TTX.

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Advanced Materials Research (Volumes 466-467)

Pages:

220-224

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https://doi.org/10.4028/www.scientific.net/AMR.466-467.220

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Online since:

February 2012

Authors:

Qing Ping Zhong, Yan Ting Liu, Yuan Ming Sun, Hoi Fu Yu, Hong Tao Lei

Keywords:

ELISA, Monoclonal Antibody, Tetrodotoxin

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