Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of 6-benzylaminopurine and its ribose adduct in bean sprouts.
Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of 6-benzylaminopurine and its ribose adduct in bean sprouts.
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DOI：
10.1016/j.foodchem.2016.03.103
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发表时间：
2016-09
期刊：
影响因子：
8.8
通讯作者：
Wei Zhang;Lishan He;Rui Zhang;Suqin Guo;Huanfang Yue;Xiangxue Ning;Guiyu Tan;Qing X. Li;Baomin Wang
中科院分区：
文献类型：
--
作者：
Wei Zhang;Lishan He;Rui Zhang;Suqin Guo;Huanfang Yue;Xiangxue Ning;Guiyu Tan;Qing X. Li;Baomin Wang
6-苄基氨基嘌呤（6-BA）是一种细胞分裂素类植物生长调节剂，在我国已被禁止用于豆芽生产。用特异性单克隆抗体（mAb 3E 5）建立了间接竞争酶联免疫吸附试验（icELISA）。该试验显示半最大抑制浓度（IC 50）和检测范围分别为18.9 ng/mL和3.6-106 ng/mL。在自制豆芽样品中6-BA的加标回收率为75%~ 89%，与液相色谱-电喷雾四极杆轨道阱质谱（LC-ESI-MS）测定结果的相关系数（R2）为0.998。LC-ESI-MS显示阳性样品中6-BA已部分代谢为6-苄氨基嘌呤核苷（6-BAR）。由于mAb 3E 5与6-BAR的交叉反应性为315%，因此icELISA测定的6-BA含量比LC-ESI-MS高5-70倍。这种对6-BAR超敏感的icELISA将允许通过检测作为潜在生物标志物的6-BAR来快速监测6-BA。
6-Benzylaminopurine (6-BA), a cytokinin plant growth regulator, has been banned for use in bean sprout production in China. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed with a specific monoclonal antibody (mAb 3E5). The assay showed a half-maximum inhibition concentration (IC50) and detection range of 18.9 ng/mL and 3.6–106 ng/mL, respectively. Recoveries of 6-BA spiked in home cultured bean sprout samples averaged from 75% to 89% with a correlation coefficient (R2) of 0.998 between the results determined by icELISA and those by liquid chromatography–electrospray ionization quadrupole Orbitrap mass spectrometry (LC–ESI-MS). LC–ESI-MS showed that 6-BA had been partially metabolized to 6-benzylaminopurine riboside (6-BAR) in the positive samples. The content of 6-BA determined by icELISA was about 5–70 times higher than that of LC–ESI-MS because mAb 3E5 had 315% cross-reactivity with 6-BAR. Such icELISA being ultra-sensitive to 6-BAR would allow quick monitoring of 6-BA by detecting 6-BAR as a potential biomarker.