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A competitive immunoassay based on engineered magnetic/fluorescent nanoparticles and biolayer interferometry-based assay for T-2 toxin determination

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Abstract

For the first time a competitive immunoassay was developed by employing T-2 antibody-functionalized magnetite nanoparticles and T-2 toxin-conjugated fluorescent quantum dots (QDs). Free T-2 and the T-2-modified QDs compete for binding to antibody-modified magnetic beads; the magnetic beads collected by magnetic separation were subjected to fluorescence intensity analysis (with excitation/emission wavelengths at 460/616 nm). This competitive immunoassay for T-2 toxin determination was applied both in a microcentrifuge tube and on a 96-well plate. The dynamic range of the immunoassay is 1–100 ng mL−1, the limit of detection (LOD) is 0.1 ng mL−1, and determination was completed in about 40 min and 30 min in the microcentrifuge tube and 96-well plate, respectively. Moreover, the biolayer interferometry (BLI) technique was employed for T-2 determination for the first time, in which the conjugate of T-2 toxin and bovine serum albumin (BSA) was immobilized on the sensors before detection. Its average recovery of T-2 toxin from barley sample ranged from 82.00 to 123.33%, and the relative standard deviation (RSD) was between 9.42 and 15.73%. The LOD of the BLI-based assay is 5 ng mL−1, and it only takes 10 min to finish the determination.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 81771965, 31571017, 81571791, 81271697), the National “Significant New Drug Creation” Science and Technology Major Program (No. 2012ZX09503001-003), the project of Science and Technology Department of Jilin Province, China (No. 20170204027NY, 20130206069GX), and the Special Project of Biological Medicine of Jilin Province, China (No. SXGJSF2017-1).

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Authors and Affiliations

  1. Key Laboratory for Molecular Enzymology and Engineering, the Ministry of Education, National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, 130012, People’s Republic of China

    Yafan Li, Li Xu, Xiaochen Fu, Hongxiang Liu & Yi Guo

  2. Institute of Biotechnology, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, People’s Republic of China

    Guo-Bin Ding

  3. Key Laboratory for Special Functional Materials, the Ministry of Education, Henan University, Kaifeng, 475004, People’s Republic of China

    Lin Song Li

  4. China-Japan Union Hospital of Jilin University, Changchun, 130033, People’s Republic of China

    Jinming Zhu

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  1. Yafan Li
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  2. Li Xu
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  3. Xiaochen Fu
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Correspondence to Li Xu or Guo-Bin Ding.

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Li, Y., Xu, L., Fu, X. et al. A competitive immunoassay based on engineered magnetic/fluorescent nanoparticles and biolayer interferometry-based assay for T-2 toxin determination. Microchim Acta 187, 514 (2020). https://doi.org/10.1007/s00604-020-04493-3

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