Vaccine Stabilization via Yeast

Methods

Summary

1. The genes for the Hepatitis B surface antigen (HBsAg) will be cloned into a shuttle vector.
2. E. Coli K12 will be transformed with the vector containing the antigen.
3. The vector will be mini-prepped from the e. coli and then sent for sequencing to ensure that the genes are in tact.
4. Then the yeast will be transformed with the vector and grown up.
5. The yeast will secrete the antigen into the media which will then be filtered from the yeast and tested for the antigen using the HBsAg ELISA kit.
6. The yeast will be desiccated using a desiccator and then re-hydrated, the ELISA test will then be performed again.

Challenges

It will take some time to ensure to that the genes are correctly assembled into the vector.  In addition to that we would need to take due care to ensure that a reasonable amount of yeast are viable after desiccation.

Pre Analysis Plan

The sequencing of the genes will determine whether or not the genes were properly inserted into a vector. The ELISA kit will be the main mode of testing whether or not the yeast is able to produce the antigen.