title: Prediction and identification of B cell epitopes derived from EWS/FLI-l fusion protein of Ewing's sarcoma.
abstract: To predict B cell epitope of Ewing's sarcoma EWS/FLI-l fusion protein and to analyze its antigenicity and immunogenicity. Comprehensive algorithms were applied to predict the possible B cell epitopes of EWS/FLI-l fusion protein. High-performance liquid chromatography (HPLC) and mass spectrometry (MS) analysis were performed to identify the synthesized epitope peptides, ELISA assays and Western blot to detect the antigenicity, and the immunogenicity of epitope peptides. Three B cell epitopes were screened out, and HPLC and MS analysis confirmed all three synthesized epitope peptides were demandable. ELISA assays verified all three epitope peptides could prime intense antigen-antibody reaction and induce ideal antibody titers after immunization to the New Zealand white rabbit. However, Western blot confirmed that antiserum of one of these epitope peptides could not recognize EWS/FLI-1 protein. Two B cell epitopes, PQDGNKPTETSQPQ and DPDEVARRWGQRKS, derived from EWS/FLI-l protein, are identified to have potential antigenicity and immunogenicity.
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, People's Republic of China.
authors.forename: Huiwen
authors.initials: H
authors.lastname: Liu
authors.forename: Lu
authors.initials: L
authors.lastname: Huang
authors.forename: Jiaquan
authors.initials: J
authors.lastname: Luo
authors.forename: Wenzhao
authors.initials: W
authors.lastname: Chen
authors.forename: Zhanmin
authors.initials: Z
authors.lastname: Zhang
authors.forename: Xiang
authors.initials: X
authors.lastname: Liao
authors.forename: Min
authors.initials: M
authors.lastname: Dai
authors.forename: Yong
authors.initials: Y
authors.lastname: Shu
authors.forename: Kai
authors.initials: K
authors.lastname: Cao
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000276:immunology; D015153:Blotting, Western; D019496:Cancer Vaccines / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000276:immunology*; D017433:Protein Structure, Secondary; D051789:Proto-Oncogene Protein c-fli-1 / Q000737:chemistry / Q000276:immunology*; D034802:RNA-Binding Protein EWS / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D012512:Sarcoma, Ewing / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; C106031:EWS-FLI fusion protein; D018985:Epitopes, B-Lymphocyte; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS