Preparation and identification of a single-chain variable fragment antibody against Newcastle diseases virus F48E9
作者： Benqiang Li , Jiaxin Ye , Yuan Lin , Man Wang , Jianguo Zhu
DOI: 10.1016/J.VETIMM.2014.08.009
关键词:
摘要: This article describes a proposed method for convenient and efficient detection of Newcastle diseases virus (NDV) that uses the fusion single-chain variable fragment (scFv) pOPE101 vector. In order to select single chain against NDV F48E9, total RNA was extracted from spleen immunized chicken, then converted into cDNA via reverse transcription. The scFv spliced by using splice-overlap extension polymerase reaction (SOE-PCR). gene cloned vector expressed in E. coli. Under optimized conditions, antibody affinity studied indirect ELISA. One positive clone selected ELISA screening, named ZL.6. Based on germline sequence, results sequence analysis showed there are more variation CDR VH VL. addition, BHK21 cell culture conducted examine potential antiviral activity experimental result demonstrated ZL.6 able neutralize F48E9 which infected cells. So will be proved useful further characterization as diagnostic tool therapeutic agent.
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