Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp.
White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV.
Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuchang, Wuhan, Hubei 430072, PR China.
Yuzhen
Y
Wang
Xiaohua
X
Zhang
Li
L
Yuan
Tao
T
Xu
Yu
Y
Rao
Jia
J
Li
Heping
H
Dai
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015686:Gene Products, env / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D016253:Microscopy, Immunoelectron; D009500:Neutralization Tests; D033561:Penaeidae / Q000821:virology*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013329:Structure-Activity Relationship; D046848:White spot syndrome virus 1
D000911:Antibodies, Monoclonal; D015686:Gene Products, env; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins