A Sensitive Immunodetection Assay Using Antibodies Specific to Staphylococcal Enterotoxin B Produced by Baculovirus Expression

• Published in: Biosensors (Basel) Vol. 12; no. 10; p. 787
• Main Authors: Jang, Ju-Hong, Kim, Sungsik, Kim, Seul-Gi, Lee, Jaemin, Lee, Dong-Gwang, Jang, Jieun, Jeong, Young-Su, Song, Dong-Hyun, Min, Jeong-Ki, Park, Jong-Gil, Lee, Moo-Seung, Han, Baek-Soo, Son, Jee-Soo, Lee, Jangwook, Lee, Nam-Kyung
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 24.09.2022; MDPI
• Subjects: Analysis; Antigens; Assaying; Baculoviridae; Baculovirus; baculovirus expression vector system; Biosensors; Care and treatment; Cell culture; Chromatography; Complications and side effects; E coli; Enterotoxins; Enzyme-linked immunosorbent assay; Flow cytometry; Immunization; immunodetection; Immunogenicity; Inflammation; Insect cells; Insects; Mass spectrometry; Mass spectroscopy; Monoclonal antibodies; Proteins; sandwich ELISA; Scientific imaging; Septic shock; Staphylococcal enterotoxin A; Staphylococcal enterotoxin B; Staphylococcus aureus infections; T cell receptors; Testing; Toxins; South Korea; United States > US
• ISSN: 2079-6374; 2079-6374
• DOI: 10.3390/bios12100787

Staphylococcal enterotoxin B (SEB) is a potent bacterial toxin that causes inflammatory stimulation and toxic shock, thus it is necessary to detect SEB in food and environmental samples. Here, we developed a sensitive immunodetection system using monoclonal antibodies (mAbs). Our study is the first to employ a baculovirus expression vector system (BEVS) to produce recombinant wild-type SEB. BEVS facilitated high-quantity and pure SEB production from suspension-cultured insect cells, and the SEB produced was characterized by mass spectrometry analysis. The SEB was stable at 4 °C for at least 2 years, maintaining its purity, and was further utilized for mouse immunization to generate mAbs. An optimal pair of mAbs non-competitive to SEB was selected for sandwich enzyme-linked immunosorbent assay-based immunodetection. The limit of detection of the immunodetection method was 0.38 ng/mL. Moreover, it displayed higher sensitivity in detecting SEB than commercially available immunodetection kits and retained detectability in various matrices and S. aureus culture supernatants. Thus, the results indicate that BEVS is useful for producing pure recombinant SEB with its natural immunogenic property in high yield, and that the developed immunodetection assay is reliable and sensitive for routine identification of SEB in various samples, including foods.