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Generation of Hybridomas Producing Monoclonal Antibodies to the NS1 Protein of the Zika Virus

机译:产生杂交瘤产生单克隆抗体的ZIKA病毒NS1蛋白

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摘要

Hybridomas producing monoclonal antibodies (mABs) to the recombinant protein NS1 of the Zika virus synthesized in human embryonic kidney HEK293 (ZV293) cells have been obtained. The specificity of antibodies has been examined by the enzyme-linked immunosorbent assay (ELISA) using the similarly synthesized NS1 proteins of flaviviruses: the Dengue 1-4 serotype virus, Japanese encephalitis virus, yellow fever virus, tick-borne encephalitis virus, and the West Nile fever virus. Two types of hybridomas whose mABs interact with thermolabile (mABs 2F6 and 3F3) and thermostable epitopes (mABs 1B9, 6D7, and 6F2) of the NS1ZV protein have been selected. It has been shown that the 3F3 mAB, which is specific to the thermolabile determinant, can be used as both binding and detecting antibodies in the sandwich ELISA. Based on the 3F3 mAB, an experimental immunochromatographic test for the detection of the NS1ZV antigen has been developed. It has also been found that mABs 1B9, 6D7, and 6F2 against thermostable epitopes of the NS1 protein specifically recognize the NS1ZV protein after its heating for 15 min to 98 degrees C.
机译:已经获得了将单克隆抗体(MAb)产生单克隆抗体(MAb)的杂交瘤已经获得于在人胚胎肾HEK293(ZV293)细胞中合成的Zika病毒的重组蛋白NS1。使用类似合成的黄病毒的NS1蛋白质的酶联免疫吸附试验(ELISA)检查了抗体的特异性:登革热1-4血清型病毒,日本脑炎病毒,黄热病病毒,蜱传脑炎病毒,以及西尼罗河热病病毒。已经选择了两种类型的杂交瘤,其MAb与热电子(MAb 2F6和3F3)和NS1ZV蛋白质的热稳定表位(MAb 1b9,6d7和6f2)相互作用。已经表明,3F3 mAb,其特异于Thermolabile决定簇,可用作夹心ELISA中的结合和检测抗体。基于3F3 mAb,已经开发了检测NS1ZV抗原的实验免疫层析试验。还发现,MAb 1b9,6d7和6f2对NS1蛋白的热稳定表位,在加热后15分钟至98℃,特别识别NS1ZV蛋白。

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