identification of a conserved b-cell epitope on reticuloendotheliosis virus envelope protein by screening a phage-displayed random peptide library守恒的b细胞表位的识别reticuloendotheliosis病毒包膜蛋白通过筛选phage-displayed随机肽库.pdfVIP

Identification of a Conserved B-cell Epitope on Reticuloendotheliosis Virus Envelope Protein by Screening a Phage-displayed Random Peptide Library 1 1 1 1 1 1 1 2 Mei Xue , Xingming Shi , Jing Zhang , Yan Zhao , Hongyu Cui , Shunlei Hu , Hongbo Gao , Xianlan Cui , Yun-Feng Wang1* 1 Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin, P. R. China, 2 Animal Health Laboratory, Department of Primary Industries, Parks, Water and Environment, Tasmania, Australia Abstract Background: The gp90 protein of avian reticuloendotheliosis-associated virus (REV-A) is an important envelope glycoprotein, which is responsible for inducing protective antibody immune responses in animals. B-cell epitopes on the gp90 protein of REV have not been well studied and reported. Methods and Results: This study describes the identification of a linear B-cell epitope on the gp90 protein by screening a phage-displayed 12-mer random peptide library with the neutralizing monoclonal antibody (mAb) A9E8 directed against the gp90. The mAb A9E8 recognized phages displaying peptides with the consensus motif SVQYHPL. Amino acid sequence of the motif exactly matched 213SVQYHPL219 of the gp90. Further identification of the displayed B cell epitope was conducted using a set of truncated peptides expressed as GST fusion proteins and the Western blot results indicated that 213SVQYHPL219 was the minimal determinant of the linear B cell epitope recognized by the mAb A9E8. Moreover, an eight amino acid peptide SVQYHPLA was proven to be the minimal unit of the epitope with the maximal bin