Hannah Spencer

Characterization of cell lines with engineered Golgi organization (NPIF)

About me

I studied my undergrad in Biochemistry at the University of York, which included a year in industry. However, I spent this year at the University of Nottingham doing an academic placement instead, which made me want to pursue a PhD. During my placement, I was tasked with writing up existing experimental results, which resulted in two publications. I returned to York and completed my final year project with Dr Daniel Ungar, working on characterizing a cell line with a mutant Golgi protein. Dani, in collaboration with Prof. Jane Thomas-Oates in York’s Centre of Excellence in Mass Spectrometry, went on to offer an NPIF PhD project to continue working with those cells. Excitingly, the project involves using the cells for the production of monoclonal antibodies in collaboration with GlaxoSmithKline. I jumped at the opportunity and now going into my third year, I’ve moved to GSK for a 6 month placement to continue working on my project in their state of the art labs.

My project

Biologics are a broad class of biopharmaceuticals ranging from monoclonal antibodies to hormones; most of which are glycoproteins. Their associated glycans can markedly alter protein properties, affecting the therapeutic efficacy of these drugs. However, current production methods are unable to generate homogenous glycoproteins due to glycan heterogeneity, arising from poorly understood glycosylation pathway variation between cell culture batches. Glycosylation has no molecular template and is instead controlled by the distribution of glycan processing enzymes in the Golgi. This is further complicated by constant turnover of Golgi cisternae, meaning resident enzymes must be recycled by retrograde trafficking.

My project involves exploring a new approach to glycosylation engineering in industrial cell lines: by targeting the interactions responsible for glycosylation enzyme sorting in the Golgi. Specifically, we are targeting the conserved oligomeric Golgi (COG) complex; a protein that co-ordinates retrograde vesicle tethering to cisternal membranes. We used CRISPR to develop a cell line with a point mutation in COG subunit 4, which impairs it’s interaction with rab30. Our aim is to characterize the effects of this mutation on production and glycosylation of glycoprotein biologics, using mass spectrometry for glycan profiling. I am currently in my third year and have just started a 6 month placement at GSK, where I’ll be using my cell lines to express some of their biologics.


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