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Development of a lateral flow dipstick immunoassay for evaluation of folate levels in maize

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Abstract

Folates (vitamin B9) are essential for all organisms as cofactors for one-carbon metabolism. However, measurement of folates is technically complicated and time-consuming. In this study, we developed a dipstick immunoassay using a folate-specific monoclonal antibody (mAb), allowing rapid and low-cost detection of folates. The indicator range of the dipstick for 5-formylterahydrofolate (5-CHO-THF), 5-methyltetrahydrofolate (5-CH3-THF) and their polyglutamyl forms was 100–200 ng mL−1; moreover, no cross-reactivity was observed with tetrahydrofolate (THF) or 5,10-methenyltetrahydrofolate (5,10-CH=THF) at 500 ng mL–1, or with the folate precursors pterin-6-COOH, p-aminobenzoate (pABA), and L-glutamate, or with the folate analogues methotrexate and 10-formyltetrahydrofolate (10-CHO-THF) at up to 1000 ng mL−1. The dipstick immunoassay was tested in maize seeds; the results classified the seeds into those with low, moderate, and high levels of folates, and were in agreement with those of liquid chromatography-mass spectrometry. Thus, we conclude that the dipstick assay will provide a versatile tool to facilitate large-scale screening of maize rich in folates.

The dipstick based immunoassay for analyzing folate level in maize

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Acknowledgements

This work was supported financially by the National Program on Key Basic Research Project of China (2013CB127003 to C.Z.), the Ministry of Science and Technology of China (2016YFD0100503 to L.J.), and the National Science Foundation of China (31400277).

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Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Qiuju Liang, Chen Yi, Ling Jiang & Chunyi Zhang

  2. College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100193, China

    Guiyu Tan & Baomin Wang

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  1. Qiuju Liang
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  2. Chen Yi
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  3. Ling Jiang
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  4. Guiyu Tan
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  5. Chunyi Zhang
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Correspondence to Guiyu Tan or Chunyi Zhang.

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Liang, Q., Yi, C., Jiang, L. et al. Development of a lateral flow dipstick immunoassay for evaluation of folate levels in maize. Anal Bioanal Chem 409, 5655–5660 (2017). https://doi.org/10.1007/s00216-017-0504-6

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  • DOI: https://doi.org/10.1007/s00216-017-0504-6

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