Abstract
Maize is one of the most important staple food crops worldwide. 5-Formyltetrahydrofolate (5-F-THF), the most dominant and stable folate derivative in maize kernels, is important to human health and acts key roles in several metabolic pathways and biological processes. To assist investigating maize germplasm and subsequent breeding of maize varieties containing high levels of 5-F-THF, a cost-effective and high-throughput methodology for 5-F-THF detection is required. In the present study, monoclonal antibodies (mAbs) against 5-F-THF were raised in mice, followed by establishment of an indirect competitive enzyme-linked immunosorbent assay (icELISA). The icELISA showed an IC50 of 35.10 ng/mL and a working range of 7.90–263.89 ng/mL for 5-F-THF. The analytical recovery for 5-F-THF in the maize kernels with icELISA was 66–111 %, and that with liquid chromatography-mass spectrometry (LC-MS) was 57–114 %, indicating a good consistency between these two methods (Slope, 0.92; R 2, 0.98). We conclude that the sensitive and versatile icELISA can be used for high-throughput detection of 5-F-THF in maize kernels.
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Acknowledgments
This work was supported by the National Program on Key Basic Research Project of China (2013CB127003) and the National Special Program for Transgenic Research (2014ZX08-003-002). The authors are also thankful to Prof. Qing X. Li for critical reading of the manuscript and for his precious suggestions.
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This study was performed strictly according to the standards described in the “Guide for the Care and Use of Laboratory Animals” (National Research Council Commission on Life Sciences, 1996 edition). All animal treatment procedures were approved by the Animal Care Committee of China Agricultural University.
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Huanfang Yue and Qiuju Liang contributed equally to this work.
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Yue, H., Liang, Q., Zhang, W. et al. A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for 5-Formyltetrahydrofolate Detection in Maize Kernels. Food Anal. Methods 9, 3155–3162 (2016). https://doi.org/10.1007/s12161-016-0503-9
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DOI: https://doi.org/10.1007/s12161-016-0503-9