基于壳聚糖微球的黄曲霉毒素B1免疫亲和柱开发制备

Development and preparation of immunoaffinity column for aflatoxin B1 based on chitosan microspheres

中文关键词:  壳聚糖微球  黄曲霉毒素B1  免疫亲和柱

英文关键词:chitosan microspheres  aflatoxin B1  immunoaffinity column

中文摘要:

      目的 基于壳聚糖微球制备用于黄曲霉毒素B1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B1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2.69 μg AFB1/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B1痕量检测前处理提供一种新的方案。

英文摘要:

      Objective To prepare an immunoaffinity column for pretreatment of aflatoxin B1 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 B1 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 2.69 μg AFB1/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 B1 trace detection.