title: Epitope mapping of the infectious hematopoietic necrosis virus glycoprotein by flow cytometry.
abstract: The glycoprotein of infectious hematopoietic necrosis virus was truncated to ten overlapping fragments. All fragments were displayed on the inner membrane of the Escherichia coli periplasm. After disruption of the outer membrane, spheroplasts that had anchored with the glycoprotein fragment were incubated with an anti-glycoprotein polyclonal antibody. Prey pairs were detected and quantitated by flow cytometry with all fragments but one, G2, reacting with the polyclonal antibody. The antigenicity of all ten fragments was analyzed using conventional methods, and epitopes were localized in all fragments, except for G2 and were consistent with FCM analysis. Antigenicity of purified glycoprotein fusion proteins was confirmed by western blotting and ELISA. This method provides a rapid, quantitative and simple strategy for identifying linear B cell epitopes of a given protein.
authors.affiliation: Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences, Harbin, 150070, People's Republic of China, 279463437@qq.com.
authors.forename: Li-Ming
authors.initials: LM
authors.lastname: Xu
authors.forename: Miao
authors.initials: M
authors.lastname: Liu
authors.forename: Jing-Zhuang
authors.initials: JZ
authors.lastname: Zhao
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Cao
authors.forename: Jia-Sheng
authors.initials: JS
authors.lastname: Yin
authors.forename: Hong-Bai
authors.initials: HB
authors.lastname: Liu
authors.forename: Tongyan
authors.initials: T
authors.lastname: Lu
mesh_terms: D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000235:genetics; D005434:Flow Cytometry; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D029142:Infectious hematopoietic necrosis virus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000939:Epitopes; D006023:Glycoproteins; D014764:Viral Proteins