Abstract
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.
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Fig. 1.
Titration of sera from 3 mice immunized with 15mer synthetic peptide by ELISA method. (A) Results of 1 week after 2nd immunization. (B) Results of 1 week after 3rd immunization.
![kjlm-28-449f1.tif](/upload/SynapseXML/0039kjlm/thumb/kjlm-28-449f1.gif)
Fig. 2.
Western blot analysis of anti-toxic shock syndrome toxin-1 reactivity of the ascites of Balb/c mouse at 7 days after intraperitoneal injection of 1E9 hybridoma.
![kjlm-28-449f2.tif](/upload/SynapseXML/0039kjlm/thumb/kjlm-28-449f2.gif)
Quantities of purified toxic shock syndrome toxin-1: (0.5, 1, 5, and 10 μg in lanes 1-4, respectively); Lane 5: molecular weight marker (Precision Plus Protein Standards; Bio-Rad).