title: Selection and characterization of single-chain recombinant antibodies against phosphoprotein of Newcastle disease virus.
abstract: Phosphoprotein (P), involved in virus RNA replication and transcription, had become a new target for the research on treating Newcastle disease virus (NDV). Here we described the cloning and expression of phosphoprotein from NDV, and then screened the anti-P antibodies from the chicken single chain fragment variable (scFv) library, which were generated from chickens immunized with the ND vaccines. As a first step, the recombinant expression vector pET28a-P was successfully constructed. In a following step, two anti-P positive scFv clones from the scFv library were selected by indirect enzyme-linked immunosorbent assay (ELISA) method. The sequence analysis of two positive clones showed that there were more variation in complementary determine region (CDR) of VH and VL, and the CDR3 in VH exhibited a significant change in amino acid number and type. In another experiment, the purified scFv antibodies used in the assay was shown to be specific for NDV-P by western blot. The results indicated that the strategy we used in this experiment proved to be convenient way for screening scFv antibody, which paved a new way for the immunization diagnosis and the exploration of integrated control of NDV.
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of 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, People's Republic of 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, People's Republic of 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, People's Republic of 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, People's Republic of China.
authors.forename: Rui
authors.initials: R
authors.lastname: Jia
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of 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; D000911:Antibodies, Monoclonal / Q000235:genetics*; D000914:Antibodies, Viral / Q000235:genetics*; D015153:Blotting, Western; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D009521:Newcastle Disease / Q000276:immunology / Q000517:prevention & control; D009522:Newcastle disease virus / Q000235:genetics / Q000276:immunology*; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D057127:Single-Chain Antibodies / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; C055859:P protein, Sendai virus; D010750:Phosphoproteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014764:Viral Proteins
keywords: Newcastle disease virus; Phosphoprotein; Single-chain variable fragment