The Generation of a Recombinant Vaccine for Bourbon Virus

Methods

Summary

I will examine the sequence of the Bourbon virus envelope protein using the antigenicity bioinformatic algorithm. This algorithm examines the basic properties of the amino acid sequence and predicts whether they will be seen by the immune system.

I have chosen the envelope protein because it is on the outside of the virus and accessible to the immune system. Most viral vaccines are based on envelope proteins.

The DNA sequence of the segments that are suggested to be highly antigenic, will be synthesized and subcloned into bacterial fusion protein expression vectors.  The fusion protein will be induced, extracted and purified. The resulting purified proteins ( mixed with adjuvant ) will be injected into mice to determine the segments with the highest antigenicity. Antigenicity will be assayed by western blot for antibodies to the original injected proteins. 

Challenges

The most likely challenge  is that none of the purified proteins will generate a significant immune response. If so, we will seek alternative vaccination  strategies, including generating fusions with TLR proteins  and  trying to use a mRNA based vaccine.

Pre Analysis Plan

We will examine the titer of the immune response to the purified envelope proteins. We suspect that high titers ( > 1/20,000)  will identify  the envelope protein vaccines that will  protect against  Bourbon virus infection. At this point,  we will seek a collaboration with an academic research lab to test the efficacy of the vaccine to  protect against Bourbon virus infection.