Description:
Reference #: 01471
The University of South Carolina is offering licensing opportunities for A Nanopore Method for Identifying Single Amino Acid in Oligopeptides
Background:
The primary sequence of a protein or a peptide is essential to its identification and function. In the area of personalized diagnosis and therapeutics, an accurate proteomic information of proteins or peptides as biomarkers can much better reflect an individual’s health status than the genomic information. Classical proteomics techniques, such as mass spectrometry (MS), are less sensitive and reproducible when detecting low abundance proteins/peptides, and are also time consuming and too expensive for medical diagnostic.
Invention Description:
The nanopore technology is a promising alternative proteomic tool for protein sequencing in point-of-care and resource-limited settings. Existing nanopore studies only achieved identification of oligopeptide in its entirety due to the lack of single amino acid distinguishability. We invented a derivatization-assisted nanopore method to differentiate single amino acids towards de novo sequencing.
Potential Applications:
The nanopore technology is a promising alternative because of its single-molecule analysis capacity and simplicity. However, while the technology is maturing in DNA detection and sequencing, it still cannot sequence protein/peptide due to the lack of single amino acid distinguishability. The project will focus on the proof-of-concept of identifying the N-terminal amino acid as the first step towards de novo protein/peptide sequencing in clinical, point-of-care, and resource-limited settings by nanopore sequencers.
Advantages and Benefits:
This approach has several unique features that distinguish it from other known research efforts. This approach is a crucial point for nanopore sensing is the effective diameter and length of the sensing region (i.e. the constriction region). In existing nanopore technologies, several amino acid residues usually engage the sensing region of the pore at the same time, leading to a joint effect to the measurement, which prevents single amino acid resolution. Second, the proposed has a efficient conjugation chemistry will be employed to in situ derivatize the N-terminal end of amino acids or peptides to control the net charge distribution, ensure the anisotropic structural feature and prolong the interaction with the lumen face of the nanopore. The nanopore biosensor detects each peptide fragment translocation at single molecular level to identify its N-terminal amino acid and length by analyzing signal amplitude, dwell time, and roughness etc. using automated algorithms. The output results can be readily input to bioinformatic analysis for sequencing. Lastly, the Nanopore devices are fabricated with precision and reproducibility suitable for clinical applications. The use of hemolysin-based nanopore allows the adoption of a large variety of well-established protocols to ensure fabrication quality. These processes produce highly consistent devices to ensure reproducible results.