基于壳聚糖微球的黄曲霉毒素B_(1)免疫亲和柱开发制备  

Development and preparation of immunoaffinity column for aflatoxin B_(1)based on chitosan microspheres

作　　者：顾一丹 马悦 陈金男 蒋欣容 曹珊珊 刘畅 孔德昭 张歆炎 GU Yi-Dan;MA Yue;CHEN Jin-Nan;JIANG Xin-Rong;CAO Shan-Shan;LIU Chang;KONG De-Zhao;ZHANG Xin-Yan(School of Grain Science and Technology,Jiangsu University of Science and Technology,Zhenjiang 212003,China;Jiangsu Provincial Engineering Research Center of Grain Bioprocessing,Zhenjiang 212003,China;Academy of National Food and Strategic Reserves Administration,Beijing 100037,China;The Quality Monitoring Center for Food and Strategic Reserves of Zhenjiang City,Zhenjiang 212003,China;College of Environmental and Chemical Engineering,Jiangsu University of Science and Technology,Zhenjiang 212003,China)

机构地区：[1]江苏科技大学粮食学院,镇江212003 [2]江苏省粮食生物加工工程研究中心,镇江212003 [3]国家粮食和物资储备局科学研究院,北京100037 [4]镇江市粮食和物资储备质量监测中心,镇江212003 [5]江苏科技大学环境与化学工程学院,镇江212003

出　　处：《食品安全质量检测学报》2024年第7期217-224,共8页Journal of Food Safety and Quality

摘　　要：目的基于壳聚糖微球制备用于黄曲霉毒素B_(1)检测前处理的免疫亲和柱。方法通过优化化学交联法的反应条件,包括壳聚糖分子量、氨基和醛基摩尔比、壳聚糖浓度和水相油相比,制备粒径大小合适且均一的壳聚糖微球,并以此为载体偶联黄曲霉毒素B_(1)的单克隆抗体制备免疫亲和柱。对亲和柱性能进行分析,使用阳性实际样品对亲和柱使用效果进行验证。结果在最佳制备条件下免疫亲和柱非特异性吸附率为5.26%,对单克隆抗体偶联率为90.90%,柱容量为2.69μg AFB_(1)/mL壳聚糖微球,检测回收率为92.12%~98.62%,相对标准偏差为0.61%~2.53%。用于实际样品检测回收率为87.90%~96.37%。结论本研究建立的免疫亲和柱制备过程简单、成本低,为黄曲霉毒素B_(1)痕量检测前处理提供一种新的方案。Objective To prepare an immunoaffinity column for pretreatment of aflatoxin B_(1)detection based on chitosan microspheres.Methods By optimizing the reaction conditions of chemical crosslinking method,including molecular weight,molar ratio of amino group to aldehyde group,concentration of chitosan and ratio of water phase-oil phase,chitosan microspheres with appropriate particle size and uniformity were prepared.Then these microspheres were coupled with monoclonal antibodies against aflatoxin B_(1)to fabricate immunoaffinity columns.The properties of affinity column were analyzed,and the effects of affinity column were validated by positive actual samples.Results Under the optimal conditions,the non-specific adsorption rate of immunoaffinity column was 5.26%,while the coupling rate of monoclonal antibody reached 90.90%.The capacity of column was 2.69μg AFB_(1)/mL chitosan microspheres,with the recovery rates ranging from 92.12%to 98.62%and the relative standard deviations between 0.61%and 2.53%.Moreover,the recoveries of the actual samples ranged from 87.90%to 96.37%.Conclusion The preparation process of the immunoaffinity column established in this study is simple and low-cost,which provides a new scheme for the pre-treatment of aflatoxin B_(1)trace detection.

关 键 词：壳聚糖微球 黄曲霉毒素B_(1) 免疫亲和柱 

分 类 号：O657[理学—分析化学] TS207.5[理学—化学]