玉米赤霉烯酮结构类似物在免疫亲和柱柱容量实时监测中的应用  

Application of zearalenone structural analogues in real-time monitoring of column capacity in immunoaffinity columns

作　　者：吴子尧 马海灵 吴光杰 赵佳莹 王李平 刘仁荣 WU Zi-Yao;MA Hai-Ling;WU Guang-Jie;ZHAO Jia-Ying;WANG Li-Ping;LIU Ren-Rong(College of Life Sciences,Jiangxi Science&Technology Normal University,Nanchang 330000,China)

机构地区：[1]江西科技师范大学生命科学学院,南昌330000

出　　处：《食品安全质量检测学报》2025年第3期25-34,共10页Journal of Food Safety and Quality

基　　金：国家自然科学基金项目(31960501);江西省教育厅科技落地计划项目(KJLD14067)。

摘　　要：目的探索一种可以在检测过程中实时监测玉米赤霉烯酮(zearalenone,ZEN)免疫亲和柱柱容量的方法。方法将两种ZEN的天然结构类似物:α-玉米赤霉烯醇(α-zearalanol,α-ZEL)、β-玉米赤霉烯醇(β-zearalanol,β-ZEL)与戊二酸酐酯化反应合成了ZEN的两种人工结构类似物(α-ZEL-G、β-ZEL-G),样品纯化后,通过荧光、紫外、高分辨率质谱法对其进行了鉴定;利用免疫亲和柱的抗体建立了竞争酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)曲线,通过半抑制浓度(half maximal inhibitory concentration,IC_(50))比较它们与抗体之间的亲和力。分别将α-ZEL-G和β-ZEL-G作为柱容量示踪物,与不同浓度的ZEN一起加入免疫亲和柱结合,探索建立一种免疫亲和柱柱容量实时监测的方法。结果ZEN、α-ZEL-G和β-ZEL-G的竞争ELISA曲线的IC_(50)分别为:2.0、1.3和10.0 ng/mL,表明α-ZEL-G与抗体的亲和力略高于ZEN,而β-ZEL-G与抗体的亲和力则明显低于ZEN与抗体的亲和力。实验结果表明:α-ZEL-G在不同柱容量的免疫亲和柱中会影响ZEN的回收率,而β-ZEL-G的加入量在柱容量50%时,不同浓度的ZEN的回收率大于80%。结论与抗体亲和力更低的β-ZEL-G更适合用于柱容量实时监测,在实际样品的检测中显示出良好的应用效果。Objective To explore a method that can monitor the immunoaffinity column capacity of zearalenone(ZEN)in real time during detection.Methods The 2 kinds of artificial structural analogues of ZEN(α-ZEL-G andβ-ZEL-G)were synthesized by esterification ofα-zearalanol(α-ZEL)andβ-zearalanol(β-ZEL)with glutaric anhydride in this study.The products were purified and identified by fluorescence,ultraviolet and high-resolution mass spectrometry.A competitive enzyme-linked immunosorbent assay(ELISA)curve was established using the same antibody with immunoaffinity columns,and their affinity with the antibody was compared by half maximal inhibitory concentration(IC_(50)).α-ZEL-G and β-ZEL-G were used as column capacity tracers and added to the immunoaffinity column together with different concentrations of ZEN to explore a method for real-time monitoring of immunoaffinity column capacity.Results The IC_(50) of the competitive ELISA curves of ZEN,α-ZEL-G andβ-ZEL-G were 2.0,1.3 and 10.0 ng/mL,respectively,indicating that the affinity ofα-ZEL-G with antibody was slightly higher than that of ZEN,while the affinity ofβ-ZEL-G with antibody was significantly lower than that of ZEN with antibody.The experimental results showed that:α-ZEL-G in immunoaffinity columns with different column sizes would affect the recovery rates of ZEN.Whereas the recovery rates of ZEN with different concentration were more than 80%while the adding concentrations ofβ-ZEL-G was 50%of the column capacity.Conclusionβ-ZEL-G with lower antibody affinity is more suitable for real-time monitoring of column volume.It shows good application results in the test of actual samples.

关 键 词：玉米赤霉烯酮 免疫亲和柱 高效液相色谱法 柱容量 实时监测 

分 类 号：TS210.7[轻工技术与工程—粮食、油脂及植物蛋白工程]