Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein.
S1D (residues 636-789) is a neutralizing epitope region on the spike protein (S) of porcine epidemic diarrhea virus (PEDV). To accurately identify epitopes on S1D, the S1-phage library containing the gene encoding the S1D region of PEDV S protein was micropanned by six specific monoclonal antibodies (McAbs) against the S1D region. These micropanned epitope regions (MER) were focused on 696-779 amino acids of the S protein. To further map epitopes of the MER, seven overlapping mini-fragments covering MER nucleotides were separately synthesized and expressed in Escherichia coli BL21 with a GST tag. These mini-GST fusion proteins were scanned by ELISA and Western blotting with the six McAbs, and the result showed that S1D5 (residues 744-759) and S1D6 (residues 756-771) are two linear epitopes of the PEDV S protein. The antisera of the epitopes S1D5 and S1D6 could react with the native S protein of PEDV. Furthermore, Pepscan of the two linear epitopes demonstrated that SS2 ((748)YSNIGVCK(755)) and SS6 ((764)LQDGQVKI(771)) are two core epitopes on S1D5 and S1D6, respectively, located on the S protein of PEDV.
Sun
Dongbo
D
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin 150001, People's Republic of China.
Feng
Li
L
Shi
Hongyan
H
Chen
Jianfei
J
Cui
Xiaochen
X
Chen
Hongyan
H
Liu
Shengwang
S
Tong
Youen
Y
Wang
Yunfeng
Y
Tong
Guangzhi
G
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D005761:Gastroenteritis, Transmissible, of Swine / Q000276:immunology* / Q000821:virology*; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000662:veterinary; D053485:Porcine epidemic diarrhea virus / Q000235:genetics / Q000276:immunology*; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D014759:Viral Envelope Proteins