Plasmonic nanopore technologies for automatic manipulation and Raman spectroscopic sequencing of single proteins

The human proteome offers unprecedented insights into health, diseases and treatment. The current protein detection techniques are either expensive, insensitive, complex or introduce biases. Meanwhile, no amplification methods for proteins exist, which complicates their detection and leads to a serious lag of proteomics behind genomics and transcriptomics, hampering not only mechanistic studies but also clinical applications. The RamanProSeq project aims to develop a label-free optical single-molecule protein sequencing technology capable of reading protein primary sequence at single-amino-acid resolution with high accuracy (>99.9%). The proposed technology will revolutionise the field of proteomics towards personalized medicine and early diagnosis of diseases. The project will be based on the partners' approaches: ultrafast Raman spectroscopy with 1 microsecond time resolution and plasmonic nanopore-enhanced Raman sensors that have demonstrated single-molecule identification of 20 proteogenic amino acids, their post-translational modifications and discrimination of single amino acid residues within single peptides. Such achievement has addressed the long-standing challenge of current single-molecule technologies that cannot detect all 20 proteinogenic amino acids. Based on these approaches, the RamanProSeq project will integrate solid-state nanopores with a high-speed Raman detection system, an automated control system, computer simulations, and advanced Raman-based bioinformatics. The output of the project will be a novel nanopore-based device that will be used to sequence individual proteins at low concentrations and with high accuracy. The project will enable low cost, high resolution and high throughput in single-protein sequencing for a broad range of applications in life science and beyond.