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Toxins 2016, 8(3), 56; doi:10.3390/toxins8030056

Quantitative Determination of Lethal Toxin Proteins in Culture Supernatant of Human Live Anthrax Vaccine Bacillus anthracis A16R

Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China
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Academic Editor: Shihui Liu
Received: 27 October 2015 / Revised: 4 February 2016 / Accepted: 5 February 2016 / Published: 25 February 2016
(This article belongs to the Collection Anthrax Toxins)
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Abstract

Bacillus anthracis (B. anthracis) is the etiological agent of anthrax affecting both humans and animals. Anthrax toxin (AT) plays a major role in pathogenesis. It includes lethal toxin (LT) and edema toxin (ET), which are formed by the combination of protective antigen (PA) and lethal factor (LF) or edema factor (EF), respectively. The currently used human anthrax vaccine in China utilizes live-attenuated B. anthracis spores (A16R; pXO1+, pXO2−) that produce anthrax toxin but cannot produce the capsule. Anthrax toxins, especially LT, have key effects on both the immunogenicity and toxicity of human anthrax vaccines. Thus, determining quantities and biological activities of LT proteins expressed by the A16R strain is meaningful. Here, we explored LT expression patterns of the A16R strain in culture conditions using another vaccine strain Sterne as a control. We developed a sandwich ELISA and cytotoxicity-based method for quantitative detection of PA and LF. Expression and degradation of LT proteins were observed in culture supernatants over time. Additionally, LT proteins expressed by the A16R and Sterne strains were found to be monomeric and showed cytotoxic activity, which may be the main reason for side effects of live anthrax vaccines. Our work facilitates the characterization of anthrax vaccines components and establishment of a quality control standard for vaccine production which may ultimately help to ensure the efficacy and safety of the human anthrax vaccine A16R. View Full-Text
Keywords: Bacillus anthracis; anthrax toxin; lethal toxin; protective antigen; lethal factor; anthrax vaccine; A16R; quantitative determination Bacillus anthracis; anthrax toxin; lethal toxin; protective antigen; lethal factor; anthrax vaccine; A16R; quantitative determination
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MDPI and ACS Style

Zai, X.; Zhang, J.; Liu, J.; Liu, J.; Li, L.; Yin, Y.; Fu, L.; Xu, J.; Chen, W. Quantitative Determination of Lethal Toxin Proteins in Culture Supernatant of Human Live Anthrax Vaccine Bacillus anthracis A16R. Toxins 2016, 8, 56.

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