Split Aptamer Ligation for Analyte Detection in Complex Fluids
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Solution Overview
Problem
Current DNA-templated reactions for molecular recognition and analyte detection rely on sequence-defined affinity, which limits their versatility and compatibility with complex biological samples.
Innovation Solution
The use of split aptamers, such as nucleic acid strands, that ligate in the presence of specific analytes, facilitating a chemical reaction like strain-promoted azide-alkyne cycloaddition, enabling dose-dependent detection of small molecules in complex fluids through lateral flow assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If DNA-templated reactions are used for molecular recognition, then detection capability is improved, but versatility is limited due to dependence on sequence-defined affinity
Solution Approach 1:
The aptamer is divided into two separate segments (first split aptamer segment and second split aptamer segment) that can independently exist but come together upon analyte binding. This segmentation allows the system to maintain high detection precision through specific aptamer-analyte interactions while gaining versatility by allowing different aptamer segments to be combined for different analytes without re designing the entire system.
Solution Approach 2:
The system transitions from static sequence-defined affinity to dynamic analyte-dependent assembly. The split aptamer segments dynamically assemble into a functional aptamer structure only when the target analyte is present, enabling the system to adapt its recognition capability based on the analyte being detected.
2Measurement precision
If traditional DNA-templated reactions are used, then nucleic acid detection is effective, but compatibility with complex biological samples is limited
Solution Approach 1:
The small molecule analyte acts as an intermediary that facilitates the assembly of split aptamer segments. This intermediary-mediated assembly mechanism allows the system to function effectively in complex biological samples by relying on the specific analyte to drive the reaction, rather than depending on general nucleic acid hybridization that can be interfered with by complex matrices.
3Measurement precision
If split aptamer ligation is implemented, then analyte-specific detection is improved, but reaction conditions must be carefully controlled
Solution Approach 1:
The split aptamer segments perform self-assembly into a functional aptamer structure when the analyte is present, without requiring external enzymes or complex reaction conditions. The analyte itself serves as the trigger for ligation, simplifying the reaction control requirements while maintaining high analyte-specific detection precision.
Data Source
AI summary
Methods, assays, and products for the detection of analytes in a sample are provided. In one aspect, for example, a device for detecting an analyte in a sample can include a fluid transfer membrane further including a sample input region operable to receive a liquid sample, a reagent region including a first split aptamer segment, a second split aptamer segment, and a detection marker, where the first and second split aptamers are operable to ligate in the presence of the analyte. The detection marker is operable to bind to the second split aptamer. The device can further include a test region having an immobilized binding reagent operable to bind to the first split aptamer segment such that the detection marker is held in the test region when the first split aptamer segment is ligated to the second split aptamer segment due to the analyte being present in the sample.