赭曲霉毒素A结构修饰物监测免疫亲和柱容量  

Monitoring the immunoaffinity column capacity by structural modification of Ochratoxin A

作　　者：侯书敏 吴珂 王李平 吴光杰[1,2] 刘仁荣 熊辉煌[3] 赵佳莹 HOU Shumin;WU Ke;WANG Liping;WU Guangjie;LIU Renrong;XIONG Huihuang;ZHAO Jiaying(School of Life Science,Jiangxi Science&Technology Normal University,Nanchang 330013,China;Jiangxi Province Key Laboratory of Natural Microbial Medicine Research,Jiangxi Science&Technology Normal University,Nanchang 330013,China;School of Chemistry and Chemical Engineering,Nanchang University,Nanchang 330031,China)

机构地区：[1]江西科技师范大学生命科学学院,江西南昌330013 [2]江西科技师范大学天然微生物药物研究江西省重点实验室,江西南昌330013 [3]南昌大学化学化工学院,江西南昌330031

出　　处：《南昌大学学报(理科版)》2025年第1期76-81,共6页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(31960501);江西省教育厅科技落地计划(KJLD14067);江西省职业早期青年科技人才培养专项项目(20244BCE52251)。

摘　　要：为探索免疫亲和柱串联高效液相色谱(immunoaffinity column-high performance liquid chromatography,IAC-HPLC)监测IAC柱容量的方法,本研究利用赭曲霉毒素A(Ochratoxin A,OTA)与乙二胺发生缩合反应,纯化获得高纯度OTA结构修饰物(OTA-X),通过光谱、质谱对其进行了鉴定,以IAC的抗体建立了酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA),测得OTA与OTA-X在竞争抑制曲线的IC50值分别为0.71和9.97 ng·mL^(-1),表明OTA-X与抗体的亲和力低于OTA。以OTA-X作为IAC柱容量的示踪物,建立了IAC-HPLC方法,监测添加不同浓度的OTA-X和OTA在IAC的回收率。结果表明:在柱容量范围内,OTA的回收率大于80%,而OTA-X的回收率随着OTA的增加而降低,总结合量大于柱容量,说明OTA-X可以作为IAC柱容量的示踪物监测柱容量,且不影响OTA的检测结果,为提高OTA检测的准确度提供了新方法。In order to monitor IAC column capacity in the immunoaffinity column-High performance liquid chromatography(IAC-HPLC)method for analyzing Ochratoxin A(OTA),OTA was reacted with ethylenediamine by a condensation reaction to acquire a structural modification OTA-X.After purification,OTA-X was identified by spectroscopy and mass spectrometry techniques.An Enzyme-Linked immunosorbent assay(ELISA)was developed based on IAC antibodies.The IC50 values for OTA and OTA-X in the competitive inhibition curves of ELISA were 0.71 and 9.97 ng·mL^(-1),respectively,indicating that the affinity for antibodies was lower than that of OTA.Using OTA-X as the tracer for assessing IAC column capacity,the recovery rates of OTA and OTA-X at different concentrations in IAC were monitored.The results showed that,within the operational column capacity range,the recovery rate for OTA remained relatively stable above 80%,while the recovery rate of OTA-X decreased with the increase of OTA and the total binding amount was above the capacity of IAC.This research showed that OTA-X can serve as a tracer for monitor IAC column capacity without affecting the accuracy of detecting OTA,providing an innovative method to enhance precision in detecting OTA.

关 键 词：免疫亲和柱 柱容量 赭曲霉毒素A 结构修饰物 高效液相色谱 

分 类 号：O657.72[理学—分析化学] TS207.3[理学—化学]