Next Generation Protein Sequencing

Multiple protein forms, called proteoforms, can be produced via alternative splicing and post-translational modifications (PTMs) to produce functionally distinct forms. Uncovering the full depth of proteoform diversity will greatly advance our understanding of human physiology and diseases. However, proteoforms typically share high amino acid sequence identity, which makes them difficult to distinguish. While current methods have improved in recent years, limitations such as lack of proteoform-specific antibodies, inherent peptide sampling bias, and limitations on throughput and analyzable mass range highlight the need for analytical alternatives.

Recently, several new approaches for single molecule protein sequencing have emerged. With single AA resolution, these technologies are promising for distinguishing proteoforms by detecting subtle AA variations and PTMs. Since these technologies are not bound by the same limitations of current approaches, they may complement current analytical approaches and enable deeper investigation into proteoform diversity and functions. Currently, our lab is most actively exploring the application of N-terminal AA recognizer (NAAR)-based sequencing and fluorosequencing for detecting peptides that can distinguish between distinct proteoforms in collaboration with Quantum SI.