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

Development of a lateral flow dipstick immunoassay for evaluation of folate levels in maize
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开发用于评估玉米叶酸水平的侧流试纸免疫测定法

DOI:
10.1007/s00216-017-0504-6
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发表时间:
2017-07
期刊:
Anal Bioanal Chem
影响因子:
--
通讯作者:
王保民
王保民
中科院分区:
其他
文献类型:
--
作者:
梁秋菊;姜凌;易尘;谭贵玉;张春义;王保民

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叶酸(维生素B 9)是一碳代谢的organisms.as辅助因子所必需的。然而,叶酸的测量在技术上是复杂且耗时的。在这项研究中,我们开发了一个试纸条免疫分析使用。叶酸特异性单克隆抗体(mAb),允许快速。和低成本的检测叶酸。5-甲酰基四氢叶酸(5-CHO-THF)、5-甲基四氢叶酸(5-CH 3-THF)及其聚谷氨酰形式的试纸指示剂范围为100-200 ng mL-1;此外,在500 ng mL-1下,未观察到与四氢叶酸(THF)或5,10-亚甲基四氢叶酸(5,10-CH=THF)或与叶酸前体蝶呤-6-COOH(对氨基苯甲酸酯)的交叉反应性。(pABA)和L-谷氨酸,或与叶酸类似物甲氨蝶呤和10-甲酰基四氢叶酸(10-CHO-THF)在高达1000 ng mL-1。用试纸条免疫测定法对玉米种子进行了检测,结果将玉米种子分为低、中、高叶酸水平的种子,并与液相色谱-质谱法的结果一致。因此,我们得出结论,试纸法将提供一种通用工具,以促进富含叶酸的玉米的大规模筛选。
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.
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