Bioelectronic Sequencing via Polymerase Current Fluctuations
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Solution Overview
Problem
Current methods for sequencing biopolymers lack efficiency in determining the sequence of biopolymers based on electrical characteristics, particularly for proteins and enzymes, which limits rapid drug screening and diagnostic applications.
Innovation Solution
A bioelectronic device is used to obtain a bioelectronic signature from current fluctuations during polymerase activity, where the duration of open periods for each dNTP monomer is distinct, allowing for accurate sequencing by analyzing the overlap of open and closed periods using machine learning techniques like Hidden-Markov Modeling and Bayesian non-parametric analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current methods are used for sequencing biopolymers, then the sequencing process can be performed, but the efficiency and speed of determining the sequence are limited
Solution Approach 1:
The patent replaces traditional mechanical/optical sequencing methods with an electrical measurement system. A polymerase enzyme is positioned between two electrodes, and nucleotide incorporation is detected through changes in electrical current. This substitution of electrical detection for mechanical/optical methods enables rapid, real-time sequencing with higher throughput and reduced time requirements.
Solution Approach 2:
The polymerase enzyme itself serves as the detection mechanism. As the polymerase incorporates nucleotides during natural enzymatic activity, it generates intrinsic electrical signals that can be measured. The system leverages the enzyme's own function to provide the detection signal, eliminating the need for separate labeling or detection steps, thereby increasing sequencing efficiency and reducing time.
2Measurement precision
If traditional sequencing methods are used, then sequence information can be obtained, but the precision and accuracy of identifying each nucleotide are limited
Solution Approach 1:
The patent focuses detection on the local electrical environment at the polymerase active site. By positioning electrodes to detect current changes specifically at the nucleotide incorporation site, the system achieves high precision in identifying individual nucleotides. The measurement is localized to the immediate vicinity of the enzymatic reaction, enabling accurate base calling without requiring complex global analysis.
Solution Approach 2:
The system detects nucleotide incorporation by monitoring changes in electrical current parameters. Each nucleotide type (A, T, C, G) produces a distinct current signal pattern when incorporated by the polymerase. By analyzing these parameter changes in the electrical signals, the system achieves high accuracy in nucleotide identification while maintaining relatively simple device architecture.
3Productivity
If label-free detection is implemented for protein function analysis, then rapid screening capability is improved, but the detection sensitivity and signal clarity may be compromised
Solution Approach 1:
The polymerase enzyme generates its own detection signal through intrinsic electrical changes during nucleotide incorporation. No external labels, fluorophores, or chemical tags are required—the enzyme's catalytic activity itself produces the measurable electrical signal. This self-service approach enables rapid, label-free detection while maintaining high sensitivity, as the electrical signals generated directly at the active site are inherently strong and specific.
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 enables accurate determination of the sequence of polynucleotides by identifying each incorporated dNTP monomer with high precision, overcoming previous limitations in sequencing biopolymers and providing a rapid, label-free detection method for enzyme function and drug screening.
Implementation Method 1
obtaining a bioelectronic signature of polymerase activity based on current fluctuations as each complementary dNTP monomer is incorporated into the template polynucleotide
Implementation Method 2
the bioelectronic device comprises a polymerase functionally coupled to at least a first electrode and a second electrode
Data Source
AI summary
The present disclosure provides devices, systems, and methods related to sequencing a biopolymer. In particular, the present disclosure provides methods of obtaining a bioelectronic signature based on current fluctuations that correspond to the activity of an enzyme-of-interest. As described herein, certain aspects of the bioelectronic signature can be used to determine the sequence of a biopolymer.