Production of Chloroplast-Derived Canine Parvovirus Vaccines Antigens

Introduction

Canine parvovirus disease is caused by infection with three mutant strains of canine parvovirus type 2 (CPV-2), CPV-2a, -2b, or -2c. This disease can cause hemorrhagic gastroenteritis and myocarditis in dogs and other canines. Canine parvovirus (CPV) remains the leading cause of death in dogs. Vaccination is the most effective measure to control the spread of infection in dogs and prevent the development of clinical CPV infection. However, immune failure following CPV vaccination can occur for different reasons, such as persistence of maternal immunity at the time of vaccination, vaccination of non-responders, and circulation of different antigenic variants of the virus. The development of plant biotechnology has made it possible to use transgenic plants as an alternative to cell culture-based subunit vaccines. Scientists have successfully expressed the non-toxic fragment of tetanus toxin in the chloroplast and induced specific antibodies against tetanus through the oral route. In addition, a protective antigen against anthrax was expressed in tobacco chloroplasts and its efficacy was shown by in vitro assays.

Fig. 1. Picture of canine parvovirus vaccine and canine parvovirus 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 the field of vaccine research and development, with a variety of mature and comprehensive platforms and technologies, to provide global customers with professional solutions for the production of chloroplast-derived canine parvovirus vaccines. Our engineers developed a subunit canine parvovirus vaccine using the antigen CPV-2a, which elicits a protective immune response against CPV.

Here, we successfully developed a well-established platform to produce transgenic tobacco chloroplasts for canine parvovirus vaccine. In addition, we developed tomato and carrot plastid transformation systems to produce canine parvovirus vaccines. Our solutions are widely used by customers in research to protect animals from parvovirus infection. Our strategy is as follows:

(1) Construction of a chloroplast transformation vector containing a linear antigenic peptide (2L21).
(2) Tobacco chloroplast transformation by particle bombardment of the vector. The aminoglycoside 3'-adenoyltransferase (Spec) gene, which confers spectinomycin resistance, is also included for selection.
(3) Analysis by PCR and Southern blot to ensure the integration of the CPV VP2 fusion gene.
(4) Western blot was used to detect the expression of CPV VP2 capsid protein in transgenic tobacco.
(5) Inoculate animals with canine parvovirus vaccine with crude leaf extract or protein-rich leaf extract from transformed plants.

Features of Our Solutions

• Tobacco can be grown where it needs to be produced, and production can be expanded according to demand.
• There are many chloroplasts in each cell of tobacco, and multiple copies of the plastid genome per cell, which can achieve high levels of protein expression.
• Using codons and select validated expression cassettes to achieve optimal yields of CPV VP2 protein.
• The 2L21 peptide can be easily fused to the KLH carrier protein, effectively protecting animals from parvovirus infection.
• Multiple genes can be co-expressed in the operon to satisfy vaccines that require multiple epitopes to function properly.

Chloroplast-transformed oral canine parvovirus vaccines are safe and effective, and have important implications for protecting animals from canine parvovirus. Lifeasible has extensive knowledge and experience in the development and production processes of chloroplast-derived canine parvovirus vaccines. 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 canine parvovirus vaccines, please contact us at any time.

References

1. Molina A, Veramendi J, Hervás-Stubbs S. (2005) Induction of neutralizing antibodies by a tobacco chloroplast-derived vaccine based on a B cell epitope from canine parvovirus[J]. Virology. 342(2): 266-275.
2. Decaro N, Buonavoglia C, Barrs VR. (2020) Canine parvovirus vaccination and immunisation failures: Are we far from disease eradication?. Vet Microbiol. 247:108760.