为研究本实验室制备的一株抗25型蓝舌病病毒(BTV-25)VP7蛋白单克隆抗体(MAb)3H7识别的B细胞抗原表位,本研究利用噬菌体展示技术对3H7识别的抗原表位进行鉴定.经过3轮淘选后进行测序,测序结果经分析比对后获得共同的短肽序列为QYHAM;将该短肽序列合成后与3H7细胞上清和腹水均能够发生特异性反应;与BTV1~24型标准阳性血清的间接ELISA结果均呈现阳性反应;进一步的序列分析表明该表位的氨基酸序列QYHAM在不同血清型的BTV株间保守,确定该MAb识别的VP7抗原表位为238QYHAM242,本研究对于BT疫苗的设计及高通量检测试剂盒的研制均具有重要意义.%To identify the B-cell epitope recognized by monoclonal antibody (MAb) of 3H7 prepared by our laboratory against protein VP7 of bluetongue virus serotype 25 (BTV-25), the epitope of BTV VP7 was screened with the 3H7 MAb by phage display technology, and the positive clones were sequenced. Sequencing results were analyzed and obtained an common encoding peptide sequence of QYHAM after three rounds panning. The peptide sequence was synthesized and reacted positively with the MAb 3H7. Indirect ELISA assay indicated that peptide reacted positively with the sera of BTV1 to BTV24 serotypes. Further sequence analysis showed that the amino acid sequence of QYHAM was conserved among different serotypes of BTV strains, and found that the MAb recognized the sequence of 238QYHAM242 in VP7 of the virus. The identification of the linear epitope would facilitate for the design of BT vaccine and the development of high-throughput detection method.
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