With the biomanufacturing field expanding, the search for an alternative biomanufacturing host has become the next step in producing recombinant proteins in a cheaper manner. Komagataella Phaffii, a strain of yeast, has been identified as a less complex, lower cost option to traditional practices using Chinese hamster ovary cells (CHO). However, K. Phaffi still presents problems in manufacturing human-safe recombinant proteins, as differences in the glycosylation post-translational modification make any recombinant proteins produced by K. Phaffi toxic for human consumption. This project will focus on modifying K. Phaffi’s glycosylation pattern to that of a human’s without affecting K. Phaffi’s recombinant protein production capacity (Ford. et. al 2026).  Previous experiments utilizing CRISPR-Cas9 to modify the K. Phaffi genome was successful in increasing recombinant protein secretion. As such, upregulating genes localized in the endoplasmic reticulum and Golgi body using CRISPR-Cas9 to increase the glycosylation factor when producing trastuzumab, an anti-breast cancer drug, is a viable strategy. We hypothesize that the upregulation of these genes as well as the glycosylation pathway will result in a shorter residency time within the endoplasmic reticulum, reducing the fraction of proteins following the native glycosylation pathway. This data gained from this project, if successful, can then be used as a foundation to alleviate similar problems in other products produced by K. Phaffi.

As a person planning to pursue graduate school, I am participating in this SuperUROP in order to gain high-level research experience in my field of interest. I have been doing research in the alternative biomanufacturing host are for almost two years now, and this program allows me to use the knowledge and experience I have gained in that time to the next steps of my project. By the end of this program, I hope to have not only gathered significant data during this program but also have increased my scientific writing skills.