| 双价猪流行性腹泻病毒特异性双峰驼源单域抗体的重组表达和特征分析 |
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| 引用本文：李 丽，杨顺利，张辛铭，蔡建平，薛慧文，尹双辉.双价猪流行性腹泻病毒特异性双峰驼源单域抗体的重组表达和特征分析[J].西北农业学报,2019,28(7):1052~1060 |
| DOI:10.7606/j.issn.1004-1389.2019.07.004 |
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| 基金项目:中国农业科学院农业科技创新工程（CAAS-ASTIP-2014-LVRI-09）。 |
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| 中文摘要:旨在构建抗猪流行性腹泻病毒（PEDV）膜蛋白（M）双峰驼源sdAb（sdAb-Mc19/29/30/37）的二价抗体，并比较研究单价和二价sdAb-Mc对温度的敏感性、pH耐受性、血清抗体竞争活性及抗原捕获活性等技术指标。研究结果证实，单价和二价sdAb-Mc在30~70 ℃和pH 2~10条件下，均可以结合M蛋白，当温度高于80 ℃和pH大于11时，其与M蛋白的结合活性显著下降。此外，研究发现，重组二价抗体的血清抗体竞争活性和抗原捕获活性强于单价的抗体。研究结果说明，重组表达的二价sdAb-Mc抗体对温度和pH耐受性没有改变，但其与靶抗原结合活性得到显著提高，为研制敏感和特异的PEDV病原学快速检测方法奠定 基础。 |
| 中文关键词:猪流行性腹泻病毒 双峰驼 二价单域抗体 |
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| Expression and Characteristic Analysis of PEDV Specific Bivalent Camel-derived Single Domain Antibodies |
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| Abstract:Compared with conventional antibodies,the single-domain antibody (sdAb),characterized by small size, high solubility and higher stability, is caused extensive concern in medical and biotechnological applications. Herein, bivalent antibodies of bactrian camel-derived sdAb (sdAb-Mc19/29/30/37) against the membrane protein (M) of porcine epidemic diarrhea virus (PEDV) were constructed, and the stability in different temperature and pH conditions, competitive capacity with polyclonal serum antibodies against M protein, and capture capacity to M protein of the monovalent and bivalent sdAb-Mcwere compared. The results confirmed that both monovalent and bivalent sdAb-Mc have excellent binding activity to M protein under the condition of 30-70 ℃ and pH 2-10. However,the binding activity was decreased significantly when the temperature higher than 80 ℃ and pH greater than 11. Furthermore, the bivalent sdAb-Mc showing stronger competitive capacity to serum antibodies and capture capacity to M protein than monovalent sdAb-Mc.The results revealed that the properties of sdAb-Mc resistance to temperature and pH condition were not influenced by constructing bivalent sdAb, but its binding activity with target antigen was significantly improved.The bivalent sdAb-Mc obtained in this study can be a powerful tool for the development of sensitive and specific PEDV detection methods. |
| keywords:Porcine epidemic diarrhea virus Bactrian camel Bivalent single domain antibody |
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