title: Neutralization Analysis of a Chicken Single-Chain Variable Fragment Derived from an Immune Antibody Library Against Infectious Bronchitis Virus.
abstract: Avian infectious bronchitis virus (IBV), which is prevalent in many countries causing severe economic loss to the poultry industry, causes infectious bronchitis (IB) in birds. Recombinant single-chain variable fragments (scFvs) have been proven to effectively inhibit many viruses, both in vitro and in vivo, and they could be a potential diagnostic and therapeutic reagent to control IB. In this study, six anti-IBV chicken scFvs, ZL.10, ZL.64, ZL.78, ZL.80, ZL.138, and ZL.256, were obtained by screening random clones from an immune antibody library. An analysis of nucleotide sequences revealed that they represented distinctive genetic sequences and greatly varied in complementarity-determining region three of the heavy chain. Neutralization tests showed that ZL.10, which bound the S1 protein in western blots, inhibited the formation of syncytia in Vero cells 48 h post IBV infection and decreased the transcriptional level of nucleoprotein mRNA to 17.2%, while the other five scFvs, including ZL.78 and ZL.256, that bound the N protein did not. In conclusion, the results suggested that specific and neutralizing chicken scFvs against IBV, which can be safe and economical antibody reagents, can be produced in vitro through prokaryotic expression.
authors.affiliation: 1 School of Agriculture and Biology, Shanghai Key Lab of Veterinary Biology, Shanghai Jiaotong University , Shanghai, People's Republic of China .|2 School of Basic Medical Sciences, Ningxia Medical University , Yinchuan, People's Republic of China .|3 Department of Internal Medicine, College of Veterinary Medicine, Northwest Agriculture and Forestry University , Yangling, People's Republic of China .
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.affiliation: 1 School of Agriculture and Biology, Shanghai Key Lab of Veterinary Biology, Shanghai Jiaotong University , Shanghai, People's Republic of China .
authors.forename: Benqiang
authors.initials: B
authors.lastname: Li
authors.affiliation: 1 School of Agriculture and Biology, Shanghai Key Lab of Veterinary Biology, Shanghai Jiaotong University , Shanghai, People's Republic of China .
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Ye
authors.affiliation: 1 School of Agriculture and Biology, Shanghai Key Lab of Veterinary Biology, Shanghai Jiaotong University , Shanghai, People's Republic of China .
authors.forename: Man
authors.initials: M
authors.lastname: Wang
authors.affiliation: 3 Department of Internal Medicine, College of Veterinary Medicine, Northwest Agriculture and Forestry University , Yangling, People's Republic of China .
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
authors.affiliation: 4 ABBIOSCI, Inc. , Seattle, Washington.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 1 School of Agriculture and Biology, Shanghai Key Lab of Veterinary Biology, Shanghai Jiaotong University , Shanghai, People's Republic of China .
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D002645:Chickens; D002522:Chlorocebus aethiops; D022801:Complementarity Determining Regions / Q000235:genetics; D005260:Female; D014644:Genetic Variation; D015726:Giant Cells / Q000187:drug effects; D001351:Infectious bronchitis virus / Q000276:immunology*; D008403:Mass Screening; D009500:Neutralization Tests; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014709:Vero Cells
chemical_list: D057134:Antibodies, Neutralizing; D022801:Complementarity Determining Regions; D057127:Single-Chain Antibodies