Production and Characterization of Anti-Staphylococcal Toxic Shock Syndrome Toxin-1 Monoclonal Antibody.
10.3343/kjlm.2008.28.6.449
- Author:
Jeong Su PARK
1
;
Jae Seok KIM
;
Jongyoun YI
;
Eui Chong KIM
Author Information
1. Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea. euichong@snu.ac.kr
- Publication Type:Original Article ; English Abstract
- Keywords:
Monoclonal antibody;
Toxic shock syndrome toxin-1;
Staphylococcus aureus
- MeSH:
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
- From:The Korean Journal of Laboratory Medicine
2008;28(6):449-456
- CountryRepublic of Korea
- Language:Korean
-
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.
