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

Jeong-Su Park; Jae-Seok Kim; Jongyoun Yi; Eui-Chong Kim

doi:10.3343/kjlm.2008.28.6.449

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Park, Kim, Yi, and Kim: Production and Characterization of Anti-Staphylococcal Toxic Shock Syndrome Toxin-1 Monoclonal Antibody

Original Article

Korean J Leg Med 2008;28(6):449-456.

Published online: 14 February 2008

DOI: https://doi.org/10.3343/kjlm.2008.28.6.449

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

Jeong-Su Park, M.D.¹, Jae-Seok Kim, M.D.², Jongyoun Yi, M.D.³, Eui-Chong Kim, M.D.^1,

¹Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea

²Department of Laboratory Medicine, Hallym University College of Medicine, Seoul, Korea

³Aerospace Medical Center, Cheongju, Korea

Corresponding author: Eui-Chong Kim, M.D. Department of Laboratory Medicine, Seoul National University College of Medicine, 28 Yeongeon-dong, Jongno-gu, Seoul 110-744, Korea Tel: +82-2-2072-3500, Fax: +82-2-747-0359 E-mail: euichong@snu.ac.kr

Received 21 July 200825 September 2008 Accepted 25 September 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 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.

Keywords: Monoclonal antibody, Toxic shock syndrome toxin-1, Staphylococcus aureus

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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.

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.

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).

Fig. 3.

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

Abbreviations: MAb, monoclonal antibody; TSST-1, toxic shock syndrome toxin-1; BSA, bovine serum albumin.

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