Expanding Proteomic Horizons with Single-Molecule Proteome Analysis

To-date, proteomic analysis has been severely limited in scale and resolution. Analyzing protein samples using an intact, single-molecule approach holds the promise to achieve improved dynamic range, reproducibility, and sensitivity in addition to enabling evaluation of protein isoforms and post-translational modifications (proteoforms) with much higher resolution than is possible today. Using the Nautilus™ Proteome Analysis Platform to interrogate immobilized, intact proteins, we aim to quantify >95% of the proteome, including proteoforms, with broad dynamic range. In one set of experiments, protein mixtures were deposited on a high-density flow cell at optically resolvable locations. We acquired PrISM (Protein Identification by Short-Epitope Mapping) data and demonstrated identification of individual protein molecules through iterative probing with our multi-affinity probes with exquisite sensitivity. In another set of experiments, we analyzed proteoforms of tau using targeted reagents capable of identifying up to 2,048 proteoforms and demonstrate robust assay performance and preliminary differences in the proteoform landscape across model systems of Alzheimer’s disease. This approach has the potential to revolutionize biomedical research by delivering both highly accurate, large-scale proteomic analyses and targeted analyses at single-molecule resolution.

 

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