Characterization of Monoclonal Antibodies Generated against <i>Norwalk virus</i> Gil Capsid Protein Expressed in <i>Escherichia coli</i>

Yoda, Tomoko; Terano, Yoshitake; Suzuki, Yasuhiko; Yamazaki, Kenji; Oishi, Isao; Utagawa, Etsuko; Shimada, Akira; Matsuura, Shiro; Nakajima, Mitsutoshi; Shibata, Teppei

doi:10.1111/j.1348-0421.2000.tb02582.x

Cited by 24 publications

(32 citation statements)

References 20 publications

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“…Expression products were analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Both strands of the insert and junction of recombinant plasmids were sequenced by using the PRISM DyeDeoxy Terminator Cycle Sequencing kit (Perkin-Elmer Applied Biosystems, Inc., Foster City, Calif.) with Trx-up and Trx-dw and were analyzed with the model 310 Genetic Analyzer (Perkin-Elmer Applied Biosystems) as described previously (11).…”

Section: Methodsmentioning

confidence: 99%

“…NV strains 36, 21, 114, 96-908, 99-8, 002, and Gifu'96 were isolated, and recombinant capsid proteins of these strains were prepared as described previously (11,12). pT7 Blue was used as a TA cloning vector (Takara Biomedicals, Kusatsu, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…We expressed a large amount of several strains of NV capsid protein in an Escherichia coli system and generated monoclonal antibodies (MAbs) against GI capsid protein (12) as well as GII capsid protein (11). Two MAbs generated against recombinant GII capsid protein (recombinant NV36 [rNV36] capsid protein) reacted to recombinant capsid proteins of both genogroups as shown in the previous study of Yoda et al (12).…”

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confidence: 99%

“…pT7 Blue was used as a TA cloning vector (Takara Biomedicals, Kusatsu, Japan). The pTrx-Fus expression vector and anti-NV36 MAbs used in this study were also prepared as described previously (11,12). Another NV strain, 00-013, was isolated from a patient in our institute.…”

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confidence: 99%

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Precise Characterization of Norovirus (Norwalk-Like Virus)-Specific Monoclonal Antibodies with Broad Reactivity

Yoda¹,

Suzuki²,

Terano

et al. 2003

J Clin Microbiol

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We have been characterizing monoclonal antibodies against Norovirus (Norwalk-like virus). In the course of our study, two monoclonal antibodies generated against Norovirus genogroup II capsid protein were found to react not only to genogroup II but also to genogroup I recombinant capsid proteins. In addition, we showed that these two monoclonal antibodies reacted to a 40-amino-acid-fragment located close to the N-terminal region of genogroup II Norovirus. Similar reactivity was observed with the equivalent region of genogroup I Norovirus. In this study, we confirmed that the epitopes of the two monoclonal antibodies existed within an 11-amino-acid peptide. To obtain an idea of the reactive ranges of the two monoclonal antibodies toward different strains of Norovirus, their reactivities were investigated using 16 types of peptide constructed according to the data in GenBank and 8 recombinant capsid proteins (7 whole capsid proteins and 1 short [80-amino-acid] protein fragment). A characteristic broad reactivity of the two monoclonal antibodies is clearly shown by the results of this study. Thus, these monoclonal antibodies could be useful tools for detecting a broad range of Norovirus strains.Norwalk virus (NV) is a member of the family Caliciviridae, genus Norovirus, formerly called Norwalk-like virus and possessing a single-stranded RNA genome. NV has been a significant cause of nonbacterial infectious gastroenteritis and foodborne diseases all over the world. The virus is highly infectious and spreads through contaminated food as well as water in food-borne disease cases. In infectious gastroenteritis cases, this virus is spread from person to person within semiclosed communities such as schools, nursing homes, and hospitals. According to national food-borne disease statistics in Japan, the number of patients involved in incidents caused by NV is likely to be large. Thus, diagnosis of NV infection is extremely important for public health, since strategies for treatment of patients and control of diseases will be carried out effectively when the causative agent is diagnosed. NV has been diagnosed using electron microscopy (EM), reverse transcription-PCR (RT-PCR), and enzyme-linked immunosorbent assay (ELISA). These methods have worked efficiently in most cases. However, due to the great diversity of nucleotide sequences throughout the whole genome of NV and the capsid protein, neither ELISA nor RT-PCR detects all types of NV (3,4,5,9). In addition, the very limited numbers of particles shed in patient fecal material makes detection by EM difficult (2, 9). In cases of NV infections with homologous strains, genomic RNA detection by RT-PCR and antigen-antibody detection by ELISA can yield excellent results (1,3,4,7,8). Regardless of its variation, NV has been divided into two large genogroups, genogroup I (GI) and genogroup II (GII).We expressed a large amount of several strains of NV capsid protein in an Escherichia coli system and generated monoclonal antibodies (MAbs) against GI capsid protein (12) as well as G...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Precise Characterization of Norovirus (Norwalk-Like Virus)-Specific Monoclonal Antibodies with Broad Reactivity

Yoda¹,

Suzuki²,

Terano

et al. 2003

J Clin Microbiol

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“…The specificities and comparative reactivities of each antibody were characterized on the 96-well ELISA plates (ASAHI TECHNO GLASS Co., Funabashi, Japan) described previously (50). The absorbance was measured at 405 nm in an ELISA microplate reader Vmax (Molecular Devices Co., Sunnyvale, Calif., U.S.A.).…”

Section: Methodsmentioning

confidence: 99%

Establishment of Highly Specific and Quantitative Immunoassay Systems for Staphylococcal Enterotoxin A, B, and C Using Newly‐Developed Monoclonal Antibodies

Sasaki

Terano

Shibata

et al. 2005

Microbiology and Immunology

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Staphylococcal enterotoxin (SE) activities remain after boiling or treating with proteases. The main symptoms such as vomiting and diarrhea, are caused by the ingestion of SEs. Among SEs, SEA has been reported to be the major and most toxic protein. A highly specific and simple assay system is required to diagnose staphylococcal food poisoning. Therefore, the development of a suitable assay system is strongly anticipated. In this study, we have established a highly specific and sensitive avidin‐biotin sandwich ELISA (ABS‐ELISA) system for SEA, SEB, and SEC1 using newly‐developed monoclonal antibodies. The linearity of these systems obtained was in the range of 0.78–25 ng/ml for each SE, and furthermore, the lower concentrations of SEs could also be detected. The recoveries of SEs from murine serum, skim milk solution, and raw milk were found to be over 90%, suggesting that our systems could detect SEs without any interventions, such as these from milk or serum proteins. We were also able to quantify SEs in 22 specimens of culture supernatants of S. aureus isolated in past occurrences. Our established system should be very useful not only in the clinical field but also in various fields of investigation because of its quantification and simplicity in detecting SEs.

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Norwalk‐Like Viruses: Detection Methodologies and Environmental Fate

Meschke

Sobsey

2003

Encyclopedia of Environmental Microbiology

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Characterization of Monoclonal Antibodies Generated against Norwalk virus Gil Capsid Protein Expressed in Escherichia coli

Cited by 24 publications

References 20 publications

Precise Characterization of Norovirus (Norwalk-Like Virus)-Specific Monoclonal Antibodies with Broad Reactivity

Precise Characterization of Norovirus (Norwalk-Like Virus)-Specific Monoclonal Antibodies with Broad Reactivity

Establishment of Highly Specific and Quantitative Immunoassay Systems for Staphylococcal Enterotoxin A, B, and C Using Newly‐Developed Monoclonal Antibodies

Norwalk‐Like Viruses: Detection Methodologies and Environmental Fate

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