摘要
以邻苯二甲酸二丁酯(DBP)为研究目标,通过重氮化法合成了人工抗原DBAP-KLH、DBAP-BSA,经紫外光谱扫描分析鉴定,证实人工抗原合成成功。动物免疫后,经细胞融合和筛选得到杂交瘤细胞,制备出抗DBP单克隆抗体。研究结果表明,单克隆抗体DBP-2C7具有较高的亲和力和灵敏度,其效价为1∶1024000,对DBP的半数抑制浓度为24.3μg/L。特异性分析显示,该抗体与半抗原的交叉反应率为28.6%,与其他邻苯二甲酸酯类化合物的交叉反应率小于1%。本研究制备了DBP人工抗原和单克隆抗体,为邻苯二甲酸二丁酯免疫测定方法的建立和应用提供了重要的实验依据和基础原料。
Artificial antigens DBAP-KLH and DBAP-BSA were synthesized through diazotization using dibutyl phthalate(DBP)and carrier proteins.The successful synthesis of these antigens was validated by ultraviolet spectral scanning.Following animal immunization,hybridoma cells were generated via cell fusion and screening,and anti-DBP monoclonal antibodies were subsequently produced.Characterization results demonstrated that the monoclonal antibody DBP-2C7 possessed high affinity and sensitivity,with a titer of 1∶1024000 and a half inhibitory concentration(IC_(50))of 24.3μg/L against DBP.Specificity assessment revealed that the antibody exhibited 28.6%cross-reactivity with the hapten,while cross-reactivity with other phthalates was less than 1%.In conclusion,this study successfully prepared DBP artificial antigens and monoclonal antibodies,thereby providing an experimental basis and essential materials for the development and application of immunoassays targeting dibutyl phthalate.
作者
刘静静
张琦
刘雨茹
张文静
李春生
LIU Jingjing;ZHANG Qi;LIU Yuru;ZHANG Wenjing;LI Chunsheng(Institute of Biology,Hebei Academy of Sciences,Shijiazhuang Hebei 050081,China;School of Pharmacy,Hebei University of Chinese Medicine,Shijiazhuang Hebei 050200,China;College of Food Science and Technology,Hebei University of Science and Technolony,Shijiazhuang Hebei 050018,China;School of Life Sciences and Food Engineering,Hebei University of Engineering,Handan Hebei 056038,China)
出处
《河北省科学院学报》
2025年第4期47-52,共6页
Journal of The Hebei Academy of Sciences
基金
河北省科学院科技计划项目(25316)。
