Charged Polymer Tether for Nanopore Peptide Orientation
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Solution Overview
Problem
Current methods for protein sequencing using recognition tunneling face challenges in drawing peptides of any charge through a nanopore in a known orientation and exerting a consistent force independent of the peptide's charge, and require high sample concentrations, complicating the analysis of mixed analytes.
Innovation Solution
A device and method involving a conjugate product with charged polymers attached to peptide fragments, where the polymeric ion carries a predetermined charge, is used to pull the peptides into a nanopore by applying an electric potential difference, ensuring consistent orientation and force, and utilizing lower analyte concentrations through deterministic delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If traditional methods are used to draw peptides through a nanopore, then the system must accommodate peptides of any charge, but the force exerted on the peptide varies with its charge, resulting in inconsistent translocation and folding issues
Solution Approach 1:
The patent introduces a charged polymer tethered to the C-terminus of the peptide as an intermediary element. This polymer acts as a mediator that experiences consistent electrophoretic force from the electric field, which then transmits this force to the peptide through the tether connection. This resolves the contradiction by decoupling the force application (on the charged polymer) from the peptide's intrinsic charge properties, ensuring reliable and consistent translocation regardless of the peptide's charge state.
2Quantity of substance
If high sample concentrations are used for protein sequencing, then sufficient analyte is available for analysis, but the analysis of mixed analytes becomes complicated and less precise
Solution Approach 1:
The patent employs deterministic delivery mechanisms that transport peptides through the nanopore in a sequential, one-at-a-time manner rather than relying on random diffusion at high concentrations. This segmentation approach allows each peptide to be individually analyzed and counted, enabling precise quantification of mixed analytes even at lower overall concentrations. The charged polymer tether ensures consistent orientation and spacing, further enhancing the ability to resolve and quantify individual components in mixtures.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the efficient sequencing of peptides of any charge in a known orientation, reducing the sample requirements and improving the quantification of mixed analytes by enabling the use of lower concentrations and deterministic transport into the nanopore.
Implementation Method 1
applying an electric potential difference between said first and second volumes of fluid... the charged polymer of the product is pulled into and optionally through the orifice by electrophoresis, thereby pulling the polymer to be analyzed into and optionally through the orifice
Implementation Method 2
the trapping of target molecules by a recognition reagent (referred to as a reader molecule) tethered to tunneling electrodes, which may be referred to as recognition tunneling (or RT)... nucleic acid bases may be read by using the electron tunneling current signals generated as the nucleo-bases pass through a tunnel gap consisting of two electrodes functionalized with reader molecules
Data Source
AI summary
Embodiments of the present disclosure are directed to methods, systems and devices, for analyzing the molecules. For example, in some embodiments, a system is provided which includes a first volume of conducting fluid, a second volume of conducting fluid, an orifice in communication with the first and second volumes of fluid, and means for applying an electric potential difference between the first and second volumes of fluid. In some such embodiments, a conjugate product is provided which includes charged polymers each having attached thereto at least one first molecule for analysis, where the product carries a predetermined charge greater than the charge on the first molecule, and upon dissolving the product in the first volume of fluid, the product is directed into the orifice.