Bioelectronic Polymerase Sequencing via Current Fluctuation Signatures
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Solution Overview
Problem
Current methods for sequencing biopolymers, such as polynucleotides, lack the ability to efficiently detect and analyze protein function without the use of labels, limiting the understanding of conformational changes and functional fluctuations.
Innovation Solution
A bioelectronic device is used to sequence polynucleotides by measuring current fluctuations as complementary nucleotidepolyphosphate monomers with distinctive charges are incorporated into the template, utilizing a polymerase functionally coupled to electrodes and obtaining a bioelectronic signature based on these fluctuations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If label-free detection methods are used to measure protein function, then the complexity of the detection system is reduced and the speed of screening is improved, but the measurement precision and ability to detect conformational changes deteriorates
Solution Approach 1:
The patent introduces an intermediary mechanism where the polymerase enzyme itself acts as a transducer, converting conformational changes during nucleotide incorporation into measurable current fluctuations. The enzyme's natural conformational transitions (open/closed states) are coupled to electrode signals, allowing label-free detection while maintaining precision through the enzyme's intrinsic functional movements.
Solution Approach 2:
The patent replaces traditional mechanical or optical labeling systems with an electrical measurement system. Instead of using fluorescent labels or mechanical probes to detect conformational changes, the invention uses electrical current fluctuations through the polymerase enzyme to monitor protein function, achieving both speed and precision simultaneously.
2Measurement precision
If traditional sequencing methods with labels are used, then the measurement precision is maintained, but the device complexity and time consumption increase
Solution Approach 1:
The patent extracts and eliminates the labeling component from traditional sequencing methods. By removing fluorescent labels, optical detectors, and associated complex instrumentation, the invention simplifies the device while maintaining sequencing accuracy through direct electrical measurement of the polymerase's conformational changes during nucleotide incorporation.
Solution Approach 2:
The polymerase enzyme serves itself as both the functional catalyst and the detection transducer. The enzyme's natural conformational changes during catalysis are directly converted into electrical signals without requiring external labels or probes, enabling the system to perform both sequencing and detection functions through the enzyme's intrinsic properties.
3Loss of time
If label-free bioelectronic detection is implemented, then the time required for sequencing is reduced and productivity is improved, but the difficulty of detecting and measuring protein function increases
Solution Approach 1:
The patent replaces difficult-to-measure mechanical conformational changes with easily detectable electrical current fluctuations. By coupling the polymerase enzyme between electrodes, the enzyme's functional movements directly modulate electrical current, converting a hard-to-measure mechanical parameter into an easily measurable electrical signal that can be detected in real-time.
Solution Approach 2:
The patent changes the measurement parameter from mechanical displacement or optical signal to electrical current. This parameter transformation allows rapid detection of polymerase activity, as electrical current fluctuations can be measured with high temporal resolution, significantly reducing sequencing time while simplifying the detection methodology.
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 method allows for rapid and label-free detection of protein function, providing new insights into conformational changes and functional fluctuations of proteins, enabling efficient sequencing of biopolymers.
Implementation Method 1
a first electrode and a second electrode separated by a gap, and a protein attached to the first and second electrodes via a linker comprising a distinctive electrical charge
Implementation Method 2
the polymerase is attached to the first and second electrodes using a linker comprising streptavidin
Data Source
AI summary
The present disclosure provides devices, systems, and methods related to sequencing a biopolymer. In particular, the present disclosure relates to methods for sequencing a polynucleotide using a bioelectronic device that obtains a bioelectronic signature (e.g., current amplitude levels) of polymerase activity based on current fluctuations as complementary nucleotidepolyphosphate monomers (e.g., having distinct charges) are incorporated into the template polynucleotide.