Multivalent Polymer-Nucleotide Conjugate for Sequencing Signal Amplification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nucleic acid sequencing methods face challenges in accurately determining nucleotide identity and sequence due to limitations in signal intensity and persistence, leading to reduced accuracy and increased sequencing time.
Innovation Solution
A multivalent binding composition comprising multiple copies of a nucleotide attached to a particle, such as a polymer or dendrimer, is used to form a binding complex with a polymerase and target nucleic acid, enhancing signal intensity and persistence through increased local nucleotide concentration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional single-nucleotide binding compositions are used, then the sequencing method is simple, but the signal intensity is insufficient leading to reduced base-calling accuracy
Solution Approach 1:
The patent combines multiple copies of the same nucleotide (e.g., 2-100 copies) onto a single particle to form a multivalent binding composition. This merging of multiple nucleotide units into one composite structure amplifies the binding signal without requiring multiple separate particles, thereby improving base-calling accuracy while maintaining practical usability.
Solution Approach 2:
The invention creates a composite particle structure that integrates multiple nucleotide copies with a particle core (such as fluorescently labeled beads or magnetic particles). This composite material approach allows the particle to exhibit enhanced binding properties and signal intensity that individual nucleotides cannot achieve alone, directly addressing the signal intensity limitation.
2Productivity
If conventional sequencing methods are used, then the equipment is simple, but the sequencing time is prolonged due to insufficient signal persistence
Solution Approach 1:
By merging multiple nucleotide copies onto a single particle, the binding event produces a stronger and more persistent signal that can be detected for longer periods. This extended signal persistence allows for faster imaging and data collection, thereby increasing sequencing speed without compromising detection reliability.
Solution Approach 2:
The invention changes the concentration parameter by presenting multiple nucleotide copies at the particle surface, which increases the local effective concentration. This parameter change enhances both signal intensity and signal persistence, enabling faster sequencing cycles while maintaining accurate base-calling.
3Length of moving object
If conventional sequencing approaches are used, then the method is straightforward, but the read length is limited due to rapid signal decay
Solution Approach 1:
The composite particle structure with multiple nucleotide copies maintains stable binding and persistent signal emission over extended periods. This prolonged signal duration enables the detection of longer nucleic acid sequences, thereby increasing read length beyond the limitations of conventional single-nucleotide approaches.
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 significantly improves base-calling accuracy, reduces sequencing time, and increases read length by stabilizing the binding complex and maintaining high signal intensity during imaging, allowing for more efficient nucleic acid sequencing.
Implementation Method 1
a multivalent binding composition comprising a polymer-nucleotide conjugate under conditions sufficient to allow a binding complex to be formed between said polymer-nucleotide conjugate and the composition of step (a), wherein the polymer-nucleotide conjugate comprises two or more copies of a nucleotide
Data Source
AI summary
Multivalent binding compositions including a particle-nucleotide conjugate having a plurality of copies of a nucleotide attached to the particle are described. The multivalent binding compositions allow one to localize detectable signals to active regions of biochemical interaction, e.g., sites of protein-protein interaction, protein-nucleic acid interaction, nucleic acid hybridization, or enzymatic reaction, and can be used to identify sites of base incorporation in elongating nucleic acid chains during polymerase reactions and to provide improved base discrimination for sequencing and array based applications.