Production of Chloroplast-Derived Cholera Vaccines Antigens

Introduction

Cholera is a disease caused by a comma-shaped, Gram-negative bacterium, Vibrio cholerae. Following ingestion of the organism by the human host, Vibrio cholerae colonizes the small intestine by penetrating the mucus layer and rapidly multiplying, and produces the enterotoxins cholera toxin causing acute watery diarrhea. This toxin consists of A and B subunits. Among them, cholera toxin B subunit (CTB) is immunogenic and can bind to the GM1 ganglioside receptor on the surface of epithelial cells. Over the past two decades, cholera has increasingly affected more countries and more people, leading to prolonged outbreaks. CTB is widely used as a candidate vaccine antigen for the treatment of cholera disease. However, cholera vaccines are still too expensive for routine use in cholera-endemic areas of developing countries, especially during cholera outbreaks. Scientists have successfully assembled synthetic CTB chloroplasts into oligomers by chloroplast engineering with the same antigens as purified natural CTB.

Fig. 1. Picture of cholera vaccine and Vibrio cholerae display.

Solutions

Our engineers are passionate about engineering chloroplasts to produce edible vaccines to help control human and animal disease. Lifeasible is an expert in vaccine development with a variety of mature and comprehensive platforms and technologies to provide global customers with professional solutions for the production of chloroplast-derived cholera vaccines. Based on our developed carrot and lettuce plastid transformation system to express cholera vaccine antigens. Our strategy is as follows:

(1) We transferred the CTB gene containing the BADH gene into the chloroplasts of lettuce, and screened the transgenic plants on the medium without antibiotic selection markers.
(2) Integrate the unmodified CTB coding sequence into the chloroplast genome to form functional oligomers.
(3) PCR and Southern blot were used to analyze whether CTB gene and BADH gene were stably integrated into the chloroplast genome.
(4) The expression of CTB protein in transgenic plants was detected by Western blot.

Features of Our Solutions

• Plants expressing bacterial and viral antigens as nuclear transgenes can trigger an immune response after oral administration of the transgenic tissue.
• Plant cells containing oral vaccines enhance vaccine activity by protecting antigens from premature digestion.
• As an efficient transmucosal carrier molecule, CTB is expressed at high levels in chloroplasts, and can synthesize fusion proteins and commercialize plant-derived vaccines.
• The expression levels of lettuce chloroplasts are similar to tobacco, and the regeneration rate is as fast as tobacco, enabling rapid vaccine production.
• Avoiding expensive purification, refrigerated shipping, and oral shipping.

The chloroplast-transformed oral cholera vaccine is safe and effective and provides protection during outbreaks of cholera disease. Lifeasible has extensive knowledge and experience in the development and production processes of the chloroplast-derived cholera vaccine. Our mission is to provide customers with comprehensive, reliable, professional solutions to accelerate your research. If you are interested in our solutions for the production of chloroplast-derived cholera vaccines, please contact us at any time.

References

1. Lopez AL, Gonzales ML, Aldaba JG, Nair GB. (2014) Killed oral cholera vaccines: history, development and implementation challenges. Ther Adv Vaccines. 2(5):123-136.
2. Suleiman A A, Ibrahim K M, Al-Shaibani A B. (2013) Production of Lettuce Edible Vaccine for Cholera Disease Using Chloroplast Genetic Engineering[J]. Journal of university of Anbar for Pure science. 7(1).