title: Anchored periplasmic expression (APEx)-based bacterial display for rapid and high-throughput screening of B cell epitopes.
abstract: We truncated the VP2 protein of infectious bursal disease virus into five fragments: V1-5. 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 VP2 fragment were incubated with an anti-VP2 polyclonal antibody (pAb). Prey pairs were detected and quantitated by flow cytometry with V1, V3, V4 and V5 fragments reacting with the pAb. The antigenicity of all five fragments was analyzed, and our results indicated that epitopes were localized in V1, V3, V4 and V5, consistent with our flow cytometry analysis. Antigenicity analysis of purified VP2 fusion proteins using Western blots confirmed this. Our method provides a rapid, quantitative and simple strategy for identifying linear B cell epitopes.
authors.affiliation: College of Life Sciences, Northeast Agricultural University, Harbin, 150030, People's Republic of China, 582084383@qq.com.
authors.forename: Mo
authors.initials: M
authors.lastname: Guo
authors.forename: Li-Ming
authors.initials: LM
authors.lastname: Xu
authors.forename: Bing
authors.initials: B
authors.lastname: Zhou
authors.forename: Jie-Chao
authors.initials: JC
authors.lastname: Yin
authors.forename: Xian-Long
authors.initials: XL
authors.lastname: Ye
authors.forename: Gui-Ping
authors.initials: GP
authors.lastname: Ren
authors.forename: De-Shan
authors.initials: DS
authors.lastname: Li
mesh_terms: D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D061505:Cell Surface Display Techniques / Q000379:methods*; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry; D057166:High-Throughput Screening Assays / Q000379:methods*; D007243:Infectious bursal disease virus / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D018985:Epitopes, B-Lymphocyte; D011994:Recombinant Proteins; C082513:VP2 protein, infectious bursal disease virus; D015678:Viral Structural Proteins