title: Identification of a Highly Conserved Epitope on Avian Influenza Virus Non-Structural Protein 1 Using a Peptide Microarray.
abstract: Avian influenza virus (AIV) non-structural protein 1 (NS1) is a multifunctional protein. It is present at high levels in infected cells and can be used for AIV detection and diagnosis. In this study, we generated monoclonal antibody (MAb) D7 against AIV NS1 protein by immunization of BALB/c mice with purified recombinant NS1 protein expressed in Escherichia coli. Isotype determination revealed that the MAb was IgG1/κ-type subclass. To identify the epitope of the MAb D7, the NS1 protein was truncated into a total of 225 15-mer peptides with 14 amino acid overlaps, which were spotted for a peptide microarray. The results revealed that the MAb D7 recognized the consensus DAPF motif. Furthermore, the AIV NS1 protein with the DAPF motif deletion was transiently expressed in 293T cells and failed to react with MAb D7. Subsequently, the DAPF motif was synthesized with an elongated GSGS linker at both the C- and N-termini. The MAb D7 reacted with the synthesized peptide both in enzyme-linked immunosorbent assay (ELISA) and dot-blot assays. From these results, we concluded that DAPF motif is the epitope of MAb D7. To our knowledge, this is the first report of a 4-mer epitope on the NS1 protein of AIV that can be recognized by MAb using a peptide microarray, which is able to simplify epitope identification, and that could serve as the basis for immune responses against avian influenza.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.|Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Jiashan
authors.initials: J
authors.lastname: Sun
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Xiurong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Xuexia
authors.initials: X
authors.lastname: Wen
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Bao
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Shi
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Qimeng
authors.initials: Q
authors.lastname: Tao
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Yongping
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Xianying
authors.initials: X
authors.lastname: Zeng
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Guobin
authors.initials: G
authors.lastname: Tian
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.
authors.forename: Shimin
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Animal Influenza Laboratory of the Ministry of Agriculture and State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Hualan
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D001717:Birds; D000939:Epitopes / Q000276:immunology / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000276:immunology* / Q000821:virology; D051379:Mice; D009975:Orthomyxoviridae / Q000276:immunology*; D040081:Protein Array Analysis; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D000939:Epitopes; D017361:Viral Nonstructural Proteins