Abstract
Proteomic mapping of cell surfaces is an invaluable tool for drug development and clinical diagnostics. This work describes a new ‘snapshot imprinting’ method designed to obtain proteomic maps of cell surfaces, with the aim of identifying cell surface markers and epitopes for diagnostic and therapeutic applications. The analysis of two cancer cell lines, HN5 and MDA-MB-468, is described herein as a proof of concept, along with the selective targeting of three identified epitopes of epidermal growth factor receptor using molecularly imprinted polymer nanoparticles. 438 proteins were identified using this technique, with 283 considered to be transmembrane or extracellular proteins. The major advantage of the molecular imprinting approach developed here is the ability to analyse cell surface proteins without tedious fractionation, affinity separation or labelling. We believe that this system of protein analysis may provide a basic molecular diagnostics toolbox for precise, personalised treatment of cancer and other diseases.
Highlights
The exposure of proteins and their spatial organisation on the cell surface is a highly complex phenomenon, influenced by protein expression, protein stability, and the micro and extracellular en vironment
Snapshot imprinting revealed a significant number of cell membrane proteins and corresponding epitopes on the surface of both cell lines that may serve as promising therapeutic targets, such as epidermal growth factor receptor (EGFR), 14–3–3 proteins, CD44 and basigin [17,18,19,20]
A total of 438 proteins were identified across these two cell lines. 241 proteins were common to both cell lines, 91 were unique to HN5 cells and 106 to MDA-MB-468 cells (Fig. 2A)
Summary
The exposure of proteins and their spatial organisation on the cell surface is a highly complex phenomenon, influenced by protein expression, protein stability, and the micro and extracellular en vironment. Used ex amples include biotinylation of surface amine residues followed by proteolysis and affinity purification of labelled extracellular peptides [4,5], ‘shaving’ approaches based on controlled proteolysis of live cells to collect surface proteins [6], and filter-aided sample pre paration (FASP) methods, compatible with both of the above [7,8]. These protocols are highly useful for drug development, for the development of immunotherapies. These peptides are not necessarily ap propriate targets for antibodies and other binding agents
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