title: Preparation and identification of a single-chain variable fragment antibody against Newcastle diseases virus F48E9.
abstract: This article describes a proposed method for convenient and efficient detection of Newcastle diseases virus (NDV) that uses the fusion of single-chain variable fragment (scFv) and pOPE101 vector. In order to select the single chain variable fragment (scFv) against NDV F48E9, the total RNA was extracted from the spleen of immunized chicken, and then was converted into cDNA via the reverse transcription. The scFv was spliced by using splice-overlap extension polymerase chain reaction (SOE-PCR). The scFv gene was cloned into a pOPE101 vector and expressed in E. coli. Under the optimized conditions, antibody affinity was studied by indirect ELISA. One positive clone was selected by ELISA screening, named ZL.6. Based on the positive clone and the germline sequence, the results of sequence analysis showed that there are more variation in CDR of VH and VL. In addition, BHK21 cell culture was conducted to examine the potential antiviral activity of ZL.6. The experimental result demonstrated that ZL.6 was able to neutralize NDV F48E9 which infected BHK21 cells. So ZL.6 will be proved useful for further characterization of NDV as potential diagnostic tool and therapeutic agent.
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Benqiang
authors.initials: B
authors.lastname: Li
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Ye
authors.affiliation: Department of Pathogenic Biology and Immunology, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Man
authors.initials: M
authors.lastname: Wang
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: zhu_jg@sjtu.edu.cn.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism*; D002645:Chickens; D003001:Cloning, Molecular; D005260:Female; D008969:Molecular Sequence Data; D009521:Newcastle Disease / Q000517:prevention & control; D009522:Newcastle disease virus / Q000145:classification* / Q000276:immunology*; D012326:RNA Splicing; D012367:RNA, Viral; D057127:Single-Chain Antibodies / Q000378:metabolism*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral; D057127:Single-Chain Antibodies; D014765:Viral Vaccines
keywords: Antiviral activity; Newcastle diseases virus; Sequence comparison; scFv