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Characterization of Monoclonal Antibodies Generated against Norwalk virus Gil Capsid Protein Expressed in Escherichia coli
Author(s) -
Yoda Tomoko,
Terano Yoshitake,
Suzuki Yasuhiko,
Yamazaki Kenji,
Oishi Isao,
Utagawa Etuko,
Shimada Akira,
Matsuura Shiro,
Nakajima Masaharu,
Shibata Tadayoshi
Publication year - 2000
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2000.tb02582.x
Subject(s) - capsid , epitope , monoclonal antibody , biology , virology , norwalk virus , recombinant dna , escherichia coli , microbiology and biotechnology , virus , epitope mapping , antigen , caliciviridae , antibody , gene , biochemistry , viral disease , norovirus , genetics
The Norwalk virus ( NV ) causes outbreaks of acute non‐bacterial gastroenteritis in humans. The virus capsid is composed of a single 60 kDa protein. The capsid protein of NV36 (genogroup II, Mexico virus type) was expressed in an Escherichia coli system and ten monoclonal antibodies (MAbs) were generated against it. The reactivity of these MAbs was characterized using enzyme‐linked immunosorbent assay (ELISA) and Western blot (WB) analysis towards 20 overlapping fragments of the NV36 capsid protein expressed in E. coli . All of the MAbs recognized sequential (continuous) epitopes on the three antigenic regions. Six of the 10 MAbs recognized fragment 2 (equivalent residues 31–70), three MAbs recognized fragment 13 (residues 361–403) and one MAb recognized fragment 7 (residues 181–220), suggesting that the N‐terminal domain (residues 1–220) may contain more antigenic epitopes than the C‐terminal domain (residues 210–548). Furthermore, two MAbs (1B4 and 1F6) reacted in WB with three purified NV strains (genogroup II) derived from patients' stool samples. It was also found that genogroup I recombinant NV96–908 (genogroup I, KY89 type) could be detected as sensitively as recombinant NV36 (genogroup II) by ELISA with a set of the MAbs produced here.