Abrin toxin(AT)and ricin toxin(RT)are highly toxic,easily accessible,and pose significant public health risks due to their potential for food and water contamination.In this study,we developed highly specific antibody...Abrin toxin(AT)and ricin toxin(RT)are highly toxic,easily accessible,and pose significant public health risks due to their potential for food and water contamination.In this study,we developed highly specific antibody pairs against AT(9D10-1B8)and RT(9G11-2B7),achieving impressive limits of detection(LODs)of 0.97 ng/mL and 0.45 ng/mL,respectively.Leveraging these antibodies,we constructed a dual-channel gold nanoparticle-based immunochromatographic assay(DC-AuNP-ICA)for the rapid,simultaneous detection of AT and RT in food and water samples.The assay demonstrated visual limits of detection(vLODs)of 5 ng/mL for AT and 2 ng/mL for RT,with calculated limits of detection(cLODs)of 3.66 ng/mL and 1.59 ng/mL,respectively.Recovery tests in spiked food and lake water samples yielded results of 93.78-105.70%,with coefficients of variation between 4.05%and 8.62%,confirming high accuracy and reproducibility.The DC-AuNP-ICA provides a reliable,on-site detection tool,offering a practical solution to mitigate the risks posed by AT and RT to public safety.展开更多
采用不同的方法纯化相思子毒素单抗,寻求一种效果好、操作简便的纯化方法。采用硫酸铵沉淀法、辛酸-硫酸铵法和硫酸铵-蛋白A亲和层析法三种方法分离纯化相思子毒素单抗,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、BCA蛋白检测...采用不同的方法纯化相思子毒素单抗,寻求一种效果好、操作简便的纯化方法。采用硫酸铵沉淀法、辛酸-硫酸铵法和硫酸铵-蛋白A亲和层析法三种方法分离纯化相思子毒素单抗,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、BCA蛋白检测试剂盒(BCA Protein Assay Kit)和双抗夹心酶联免疫吸附试验(ELISA)对纯化产物进行相对分子质量、浓度、纯度、回收率及免疫活性的鉴定。结果表明,纯化后单抗的回收率以硫酸铵沉淀法及辛酸-硫酸铵法较高,硫酸铵-蛋白A亲和层析法较低;产物纯度以硫酸铵-蛋白A亲和层析法和辛酸-硫酸铵法较高,硫酸铵沉淀法较差;不同方法纯化后的单抗活性未见降低。展开更多
基金supported by the Natural Science Foundation of Jiangsu Province,China(BK20230043,BK2024304,BK20221088,BK20221058,CX(22)1013,KJ2025008,M20221006)the Major Science and Technology Projects,China(110202201020,LS-04)+1 种基金National Key R&D Program,China(2022YFF1101002)National Natural Science Foundation of China(22476067,22404066,22361132536,22306074,22236002,22207045).
文摘Abrin toxin(AT)and ricin toxin(RT)are highly toxic,easily accessible,and pose significant public health risks due to their potential for food and water contamination.In this study,we developed highly specific antibody pairs against AT(9D10-1B8)and RT(9G11-2B7),achieving impressive limits of detection(LODs)of 0.97 ng/mL and 0.45 ng/mL,respectively.Leveraging these antibodies,we constructed a dual-channel gold nanoparticle-based immunochromatographic assay(DC-AuNP-ICA)for the rapid,simultaneous detection of AT and RT in food and water samples.The assay demonstrated visual limits of detection(vLODs)of 5 ng/mL for AT and 2 ng/mL for RT,with calculated limits of detection(cLODs)of 3.66 ng/mL and 1.59 ng/mL,respectively.Recovery tests in spiked food and lake water samples yielded results of 93.78-105.70%,with coefficients of variation between 4.05%and 8.62%,confirming high accuracy and reproducibility.The DC-AuNP-ICA provides a reliable,on-site detection tool,offering a practical solution to mitigate the risks posed by AT and RT to public safety.
文摘采用不同的方法纯化相思子毒素单抗,寻求一种效果好、操作简便的纯化方法。采用硫酸铵沉淀法、辛酸-硫酸铵法和硫酸铵-蛋白A亲和层析法三种方法分离纯化相思子毒素单抗,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、BCA蛋白检测试剂盒(BCA Protein Assay Kit)和双抗夹心酶联免疫吸附试验(ELISA)对纯化产物进行相对分子质量、浓度、纯度、回收率及免疫活性的鉴定。结果表明,纯化后单抗的回收率以硫酸铵沉淀法及辛酸-硫酸铵法较高,硫酸铵-蛋白A亲和层析法较低;产物纯度以硫酸铵-蛋白A亲和层析法和辛酸-硫酸铵法较高,硫酸铵沉淀法较差;不同方法纯化后的单抗活性未见降低。
