Monoclonal Antibodies Against Nsp2 Protein of the Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
Monoclonal Antibodies Against Nsp2 Protein of the Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
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DOI：
10.1089/mab.2013.0023
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发表时间：
2013-10-01
影响因子：
--
通讯作者：
He, Kongwang
中科院分区：
文献类型：
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作者：
Li, Bin;Du, Luping;He, Kongwang
猪繁殖与呼吸综合征（PRRS）仍然是一个严重的威胁，对世界范围内的养猪业造成重大的经济影响。本研究在大肠杆菌中表达了猪繁殖与呼吸综合征病毒（PRRSV）的非结构蛋白Nsp2，并通过透析纯化。通过将小鼠骨髓瘤细胞系SP2/0与Nsp2蛋白免疫小鼠的脾脏淋巴细胞融合，制备了一种重要的抗Nsp2蛋白单克隆抗体（MAb 2H6）。然后用酶联免疫吸附试验（ELISA）、Western blot分析和间接免疫荧光法检测单抗的活性。结果表明，单抗在1:100稀释的IFA和1:500稀释的Western blot分析中对HP-PRRSV呈阳性反应，与经典PRRSV无反应。这表明该抗PRRSV Nsp2蛋白的单抗可能是一种特异性诊断方法和Nsp2蛋白功能探索的良好候选。
Porcine reproductive and respiratory syndrome (PRRS) continues to be a serious threat, causing an economically significant impact on the swine industry worldwide. In this study, non-structural protein Nsp2 of porcine reproductive and respiratory syndrome virus (PRRSV) was expressed in Escherichia coli and purified by dialysis. An important monoclonal antibody (MAb 2H6) against Nsp2 protein was generated by fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from Nsp2 protein immunized mice. Then activity of the MAb was characterized by enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and indirect immunofluorescence assays. The results demonstrated that the MAb has a positive reaction to HP-PRRSV in IFA at 1:100 dilution and in Western blot analysis at 1:500 dilution, and no reaction with classic PRRSV. These indicated that this MAb against Nsp2 protein of PRRSV might be a good candidate for a specific diagnostic method and functional exploration of the Nsp2 protein.