Production of therapeutic proteins
Another component of our research program involves the production of BBB-targeting antibodies both as therapeutic proteins, and as fusions to fluorescent proteins for use in a diagnostic setting. Production of antibodies, and in particular, antibody fragments, has been a difficult challenge. We employed yeast to produce significant quantities of antibodies while using cellular and protein engineering strategies to optimize the process. We have used our methods to significantly improve the production of an anti-transferrin receptor BBB-targeting antibody that could only be produced previously as an insoluble aggregated using bacterial production approaches. In addition, we have developed novel screening strategies that have allowed us to mine the entire yeast genome for gene products that can elevate the level of antibody secretion from yeast. We identified three yeast proteins, Ccw12p, Ero1p, and Rpp0p, that were heretofore unknown protein secretion enhancers that elevate antibody production 2-8 fold.
- Goulatis LI et al. (2019). Impacts of the -1 amino acid on yeast production of protein-intein fusions. Biotechnol Prog 35:e2736.
- Umlauf BJ et al. (2018). Site-specific antibody functionalization using tetrazine-styrene cycloaddition. Bioconjug Chem 29:1605-13.
- Burns ML et al. (2016). Pro-region engineering for improved yeast display and secretion of brain derived neurotrophic factor. Biotechnol J 11:425-36.