Mapping of epitopes of VP2 protein of chicken anemia virus using monoclonal antibodies.
To map the epitopes of VP2 protein of chicken anemia virus (CAV), VP2 was expressed as a fusion protein in Escherichia coli BL21 (DE3). The Western blot demonstrated that recombinant VP2 protein could be recognized by sera of chickens infected with CAV. Female BALB/c mice were immunized with purified recombinant VP2 produced in E. coli BL21 (DE3) and seven VP2-specific monoclonal antibodies (MAbs) were developed. The results of Western blot showed that all the seven MAbs recognized the recombinant VP2 protein expressed in the baculovirus and reacted with MDCC-MSB1 cells infected with CAV by indirect immunofluorescence assay. The VP2 protein was dissected into 21 overlapping fragments, expressed as fusion peptides in E. coli and used for epitope mapping by pepscan analysis. ELISA and Western blot assays indicated that most of MAbs reacted with the 12th and 13th fragments (amino acids 111-136) and one of them reacted with the 3rd fragment (amino acids 21-36). The linear immunodominant epitope of VP2 was located mainly in amino acid residues 111-126 and 121-136.
Wang
Xiaoyan
X
Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, 427 Maduan Street, Harbin 150001, China. xmw@hvri.ac.cn
Gao
Honglei
H
Gao
Yulong
Y
Fu
Chaoyang
C
Wang
Zhi
Z
Lu
Guili
G
Cheng
Yu
Y
Wang
Xiaomei
X
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology*; D045744:Cell Line, Tumor; D017727:Chicken anemia virus / Q000276:immunology*; D018604:Epitope Mapping; D005260:Female; D016056:Immunodominant Epitopes; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D016056:Immunodominant Epitopes; D011993:Recombinant Fusion Proteins; C442521:VP2 protein, Chicken anemia virus