Characterization of cell lines with engineered Golgi organization
Lead Research Organisation:
University of York
Department Name: Biology
Abstract
Glycans are carbohydrate polymers which, with proteins, DNA and RNA, are one of the four biopolymers essential for life. Yet our understanding of glycans remains rudimentary compared with that of proteins and nucleotides. This is a significant drawback given the recent interest in therapeutic proteins known as biologics. Most of these are glycoproteins, and their attached glycans markedly alter efficacy of these drugs. However, current production methods are unable to generate homogenous glycoproteins due to glycan heterogeneity. This project will use our knowledge of glycan biosynthesis in the Golgi apparatus to develop and characterize engineered cell lines with more favorable glycosylation properties. Cell biological and mass spectrometric methods will then be used to characterize both the organization of glycosylation enzymes in the Golgi and the glycans produced by the cells. In collaboration with GSK, the student will then have the opportunity to test the production and glycosylation of biologics using the newly developed cell lines. The student will be part of a vibrant glycobiology community and will be trained in modern cell biology (cell culture, fluorescent imaging) and mass spectrometry (glycan profiling, LC-MS, tandem MS) as well as routine molecular biology and biochemistry methods. The studentship bursary will be enhanced appropriate for a CASE collaboration and involve a 3-6 month placement at GSK, Stevenage
| Description | We are interested in exploring how living cells that make protein-based pharmaceuticals (biologics) modify those proteins with the addition of sugar molecules, a process called glycosylation. Our aim is to make mutations in the cells that might alter glycosylation and improve the production of glycosylated biologics. We are specifically targeting a protein complex, called the conserved oligomeric Golgi (COG) complex, which is responsible for organizing glycosylation enzymes within the Golgi. We have confirmed that a mutation in subunit 4 (COG4) of this protein impairs it's interaction with rab30, a GTPase protein, which is also involved in glycosylation enzyme sorting. Although we are yet to confirm whether we have achieve dour aim of altering glycosylation, initial results suggest that the mutation causes an increase in biologic production, which could have great implications for industrial production of biologics. |
| Exploitation Route | Our cell lines could be used by the pharmaceutical industry for testing the production of biologics. |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |