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A Sensitive Immunodetection Assay Using Antibodies Specific to Staphylococcal Enterotoxin B Produced by Baculovirus
Ju-Hong Jang1,2, Sungsik Kim1, Seul-Gi Kim1,2
1Biotherapeutics Translational Research Center, Korea Research Institute of Bioscience and Biotechnology, 125 Gwahak-ro, Yuseong-gu, Daejeon 34141, Korea.
Biosensors
|October 27, 2022
Summary
A new immunodetection system effectively identifies Staphylococcal enterotoxin B (SEB), a dangerous toxin. This method uses a baculovirus expression vector system (BEVS) for high-purity SEB production and sensitive monoclonal antibody detection.
Area of Science:
- Microbiology
- Immunology
- Biotechnology
Background:
- Staphylococcal enterotoxin B (SEB) is a potent bacterial toxin causing inflammation and toxic shock.
- Accurate detection of SEB in food and environmental samples is crucial for public health.
Purpose of the Study:
- To develop a sensitive immunodetection system for SEB.
- To produce recombinant wild-type SEB using a baculovirus expression vector system (BEVS).
Main Methods:
- Utilized BEVS for high-yield, pure recombinant SEB production in insect cells.
- Characterized SEB using mass spectrometry.
- Generated monoclonal antibodies (mAbs) against SEB via mouse immunization.
- Developed a sandwich enzyme-linked immunosorbent assay (ELISA) using a selected mAb pair.
Main Results:
- Achieved high-quantity and purity of recombinant SEB via BEVS.
- The produced SEB was stable for over 2 years at 4 °C.
- The developed immunodetection assay demonstrated a limit of detection of 0.38 ng/mL.
- The assay showed higher sensitivity than commercial kits and detected SEB in various matrices and bacterial supernatants.
Conclusions:
- BEVS is effective for producing pure, immunogenic recombinant SEB in high yields.
- The developed immunodetection assay is reliable and sensitive for routine SEB identification in diverse samples, including food.
