基于TLH重组蛋白单抗的磁性免疫层析试纸条构建

Establishment of magnetic immunochromatographic test strip based on monoclonal antibody against recombinant protein TLH

为了检测不耐热溶血毒素TLH,以His-TLH重组蛋白为抗原免疫BALB/c小鼠,利用杂交瘤技术制备TLH单克隆抗体,然后将纯化后的单抗与磁珠偶联制备免疫磁珠,以此免疫磁珠为标记物构建了TLH重组蛋白的磁性免疫层析检测试纸条。结果表明,该磁性免疫层析试纸条实现了TLH重组蛋白的快速定性和定量检测,具有快速、操作简便、灵敏度高(检测限12.5 ng/mL)等特点。本研究所构建的磁性免疫层析试纸条为重组溶血毒素的快速鉴定、诊断和检测提供了一种新途径。

In order to detect thermolabile hemolysin（TLH）,the recombinant protein His-TLH was used to immunize a BALB/c mouse,the monoclonal antibody（MAb） against recombinant protein TLH was prepared by hybridoma technique,and then the purified MAb was conjugated with magnetic beads to prepare immunomagnetic beads（IMBs）,thus a magnetic immunochromatographic test strip（MITS） being established by taking the IMBs as labels for His-TLH.The test result showed that the MITS could be used for qualitative and quantitative assay of recombinant protein TLH,having such advantages as simplicity,rapidity and high sensitivity（the detection limit;12.5 ng/mL）.The developed MITS in this study also provides a new means of rapid identification,diagnosis and detection of recombination hemolysin.