农科院特产所研制出犬瘟热病毒单链抗体

序道

      近日，中国农业科学院特产研究所特种动物病原与免疫创新团队研制出犬瘟热病毒单链抗体，相关研究成果发表在最新一期的国际专业学术期刊《单克隆抗体的免疫诊断和免疫治疗(Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)》上。


      据悉，犬瘟热病毒(CDV)是危害毛皮动物产业的主要疫病，目前尚无相关治疗方法，而抗体(单抗)疗法被誉为最有前景的一个研究方向。单链抗体(scFv)是一种微小的基因工程抗体，它包含抗体重链和轻链的可变区和一段链接肽。目前来说，单链抗体是动物体内清除病毒的强有力的治疗型工具，也能作为一种非常经济而且容易获得的诊断试剂组件。

      特种动物病原与免疫团队采用分泌抗犬瘟热病毒核蛋白的杂交瘤细胞RNA为模板，分离出其抗体轻链和重链可变区，通过基因工程的方式组装成单链抗体表达载体。通过原核表达，获得了杂交瘤细胞的重组单链抗体蛋白，经过间接酶联免疫吸附实验(ELISA)和竞争ELISA方法检测，其具备天然单抗的部分活性。换句话说，这种重组单链抗体蛋白能识别犬瘟热病毒并能阻断天然单抗与犬瘟热病毒核蛋白结合能力。本研究为未来研究犬瘟热病毒的新型诊断试剂和治疗试剂奠定了基础，具有很好的开发前景。


      特种动物病原与免疫团队多年来一直从事特种动物病毒分子生物学、分子免疫学等相关研究，该成果不仅填补了国内该领域的空白，而且为特种动物疫病防治提供了免疫学生物工程制剂。


      单链抗体是一种微小的基因工程抗体，是动物体内清除病毒的强有力的治疗型工具，也能作为一种非常经济而且容易获得的诊断试剂组件。


文章信息：Li Yi and Shipeng Cheng
Institute of Special Wild Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, P.R. China.
Address correspondence to:
Shipeng Cheng
Institute of Special Wild Economic Animal and Plant Science
Chinese Academy of Agricultural Sciences4899 Juye Street
Changchun
Jilin Province
P.R.China 130122
E-mail: tcscsp@126.com

Received: January 26, 2015
Accepted: April 9, 2015


原文摘要：

The variable regions of the heavy chain (VH) and light chain (VL) were amplified by RT-PCR from the hybridoma 1N8, which secretes the monoclonal antibody against CDV N protein (aa 277-471). The VL and VH amplicons were combined using SOE-PCR by a 12 amino acid flexible linker (SSGGGGSGGGGS), which produced the scFv gene (named scFv/1N8). After sequence analysis, the scFv/1N8 gene was cloned into the prokaryotic expression vector PET32a with a His-tag. The recombinant scFv/1N8 protein was successfully expressed in recombinant Escherichia coli by IPTG induction. Moreover, the binding activity and specificity of the scFv were determined by indirect ELISA (His-tag) and competitive ELISA. The recombinant scFv/1N8 protein reported here will provide some basis for further antiviral drug research based on the scFv molecule.

相关链接：http://online.liebertpub.com/doi/abs/10.1089/mab.2015.0008?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed