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KMID : 0981820080280060449
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Korean Journal of Laboratory Medicine
2008 Volume.28 No. 6 p.449 ~ p.456
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Production and Characterization of Anti-Staphylococcal Toxic Shock Syndrome Toxin-1 Monoclonal Antibody
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Park Jeong-Su
Kim Eui-Chong
Yi Jong-Youn
Kim Jae-Seok
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Abstract
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Background:Recently the association between the virulence factors of Staphylococcus aureus and the outcome of the patients infected with the organism appears to be the subject of active investigation. Toxic shock syndrome toxin-1 (TSST-1) is thought to be a clinically more significant virulence factor than other staphylococcal toxins. We attempted to produce and characterize monoclonal antibodies to staphylococcal TSST-1.
Methods:An important epitope of TSST-1, amino acids 1-15 region, was synthesized into a peptide antigen, and Balb/c mice were immunized by intraperitoneal injection of the synthetic antigen. Hybridomas were produced by fusing immunized murine splenocytes with immortal myeloma cells. Hybridomas were cloned through a limiting dilution method. Stable cultured hybridoma was injected into the peritoneal cavity of Balb/c mice, and peritoneal fluid containing the monoclonal antibody was produced.
Results:One IgG2b type monoclonal antibody and two IgM type monoclonal antibodies were obtained. The IgG2b type monoclonal antibody was able to detect 5 μg of TSST-1 with Western blot analysis and showed a strong reactivity to TSST-1 with ELISA.
Conclusions:Highly immunoreactive anti-TSST-1 monoclonal antibody was produced by the use of synthesized peptide antigen. Diagnostic and protective capacity of this monoclonal antibody should be evaluated in the future.
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KEYWORD
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Monoclonal antibody, Toxic shock syndrome toxin-1, Staphylococcus aureus
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