title: Putative phage-display epitopes of the porcine epidemic diarrhea virus S1 protein and their anti-viral activity.
abstract: Porcine epidemic diarrhea virus (PEDV) is a pathogen of swine that causes severe diarrhea and dehydration resulting in substantial morbidity and mortality in newborn piglets. Phage display is a technique with wide application, in particular, the identification of key antigen epitopes for the development of therapeutic and diagnostic reagents and vaccines. To identify antigen epitopes with specificity for PEDV, a monoclonal antibody (MAb-5E12) against the immunodominant region of the PEDV Spike protein (S1) was used as the target for biopanning a 12-mer phage display, random peptide library. After multiple rounds of biopanning and stringent washing, three phage-displayed peptides, designated L, W and H, were identified that recognize MAb-5E12. Sequence analysis showed that the one or more of the peptides exhibited partial sequence similarity to the native S1 sequence 'MQYVYTPTYYML' (designated peptide M) at position 201-212. In combination with software analysis for the prediction of B cell epitopes, aa 201-212 exhibited characteristics of a linear epitope on the PEDV S1 protein. In contrast to peptide M, a consensus motif 'PxxY' was identified on both peptides L and W, and on the S1 protein, but not on peptide H. Peptide M and the MAb-5E12-recognizing peptides L and W significantly inhibited the adsorption of PEDV on the cell surface as monitored through plaque-reduction assays. Furthermore, data from real-time PCR and indirect immunofluorescence assays were consistent with the ability of peptides M, L and W to block viral protein expression and thereby function as antiviral agents for PEDV.
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Liyan
authors: L
authors: Cao
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Xuying
authors: X
authors: Ge
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Yu
authors: Y
authors: Gao
authors: Animal Parasitic Diseases Laboratory, Agricultural Research Service, Beltsville, MD, 20705, USA.
authors: Dante S
authors: DS
authors: Zarlenga
authors: Harbin National Engineering Research Center of Veterinary Biologics Co., Ltd., 789 Chuangxin Street, Songbei District, Harbin, 150001, China.
authors: Kexiong
authors: K
authors: Wang
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Xunliang
authors: X
authors: Li
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Zhaoheng
authors: Z
authors: Qin
authors: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Epizootic Disease of Grazing Animal of Ministry of Agriculture, Lanzhou Veterinary Research Institute, 11 Yanchang Street, Chengguan District, Lanzhou, 730046, China.
authors: Xiangping
authors: X
authors: Yin
authors: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Epizootic Disease of Grazing Animal of Ministry of Agriculture, Lanzhou Veterinary Research Institute, 11 Yanchang Street, Chengguan District, Lanzhou, 730046, China.
authors: Jisheng
authors: J
authors: Liu
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China.
authors: Xiaofeng
authors: X
authors: Ren
authors: College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin, 150030, China. ligx@neau.edu.cn.
authors: Guangxin
authors: G
authors: Li
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002522:Chlorocebus aethiops; D000939:Epitopes / Q000276:immunology*; D019084:Fluorescent Antibody Technique, Indirect; D009500:Neutralization Tests; D019151:Peptide Library; D053485:Porcine epidemic diarrhea virus / Q000276:immunology* / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D014709:Vero Cells; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000276:immunology*; D053585:Virus Attachment / Q000187:drug effects
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D019151:Peptide Library; D014764:Viral Proteins
keywords: Monoclonal antibody; Phage display technique; Porcine epidemic diarrhea virus (PEDV); Putative epitope; S1 protein