Korean J Lab Med.  2008 Dec;28(6):449-456. 10.3343/kjlm.2008.28.6.449.

Production and Characterization of Anti-Staphylococcal Toxic Shock Syndrome Toxin-1 Monoclonal Antibody

• Park JS
• Kim JS
• Yi J
• Kim EC

• KMID: 1781580
• DOI: http://doi.org/10.3343/kjlm.2008.28.6.449

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. RESULTS: One IgG2b type monoclonal antibody and two IgM type monoclonal antibodies were obtained. The IgG2b type monoclonal antibody was able to detect 5 microgram 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.

Keyword

Monoclonal antibody; Toxic shock syndrome toxin-1; Staphylococcus aureus

MeSH Terms

Amino Acid Sequence
Animals
Antibodies, Monoclonal/biosynthesis/*immunology/isolation & purification
Bacterial Toxins/*immunology
Blotting, Western
Enterotoxins/*immunology
Enzyme-Linked Immunosorbent Assay
Hybridomas/metabolism
Mice
Molecular Sequence Data
Peptides/chemical synthesis/pharmacology
Superantigens/*immunology

Figure

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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.

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  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.

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  Fig. 3. Relative binding affinity of 1E9 monoclonal antibody and control IgG for toxic shock syndrome toxin-1 and bovine serum albumin by ELISA.

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