建立一种基于单克隆抗体的酶联免疫吸附分析法，用于豆芽中 6-苄基氨基嘌呤及其核糖加合物的分析。
Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of 6-benzylaminopurine and its ribose adduct in bean sprouts.
机构信息
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
出版信息
Food Chem. 2016 Sep 15;207:233-8. doi: 10.1016/j.foodchem.2016.03.103. Epub 2016 Mar 30.
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 (R(2)) 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.
6-苄氨基嘌呤（6-BA）是一种细胞分裂素植物生长调节剂，已在中国被禁止用于豆芽生产。本研究利用特异性单克隆抗体（mAb 3E5）建立了间接竞争酶联免疫吸附法（icELISA）。该方法的半抑制浓度（IC50）和检测范围分别为 18.9ng/mL 和 3.6-106ng/mL。在家庭培养的豆芽样品中添加 6-BA 的回收率平均为 75%-89%，icELISA 法与液相色谱-电喷雾-四极杆轨道阱质谱法（LC-ESI-MS）法测定结果的相关系数（R2）为 0.998。LC-ESI-MS 结果表明，阳性样品中 6-BA 已部分代谢为 6-苄氨基嘌呤核苷（6-BAR）。icELISA 法测定的 6-BA 含量约为 LC-ESI-MS 法的 5-70 倍，因为 mAb 3E5 与 6-BAR 的交叉反应率为 315%。icELISA 对 6-BAR 的超高灵敏度可以通过检测 6-BAR 作为潜在的生物标志物来快速监测 6-BA。
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