AGE-SELEX Aptamer Selection Bioactivity Resolution
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current aptamer selection methods, such as SELEX, rely solely on binding affinity and suffer from low resolution, unspecific elution, and limited yield, failing to effectively identify aptamers with specific bioactivity for in vitro or in vivo applications.
Innovation Solution
The AGE-SELEX method enhances aptamer selection by incorporating bioactivity-guided selection rounds, using mutational PCR and target variants to refine affinity and specificity, and includes bioactivity determination steps to characterize aptamers' ability to modulate biochemical or biophysical reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If SELEX method is used for aptamer selection, then binding affinity can be obtained, but resolution is low and unspecific elution occurs
Solution Approach 1:
The patent segments the selection process into distinct stages: initial SELEX for binding affinity, followed by bioactivity-guided selection rounds. This segmentation allows each stage to optimize for its specific purpose, improving overall resolution and reducing unspecific elution by separating binding selection from functional validation.
Solution Approach 2:
The patent implements feedback mechanisms by using bioactivity determination results to guide subsequent selection rounds. Aptamers showing desired bioactivity are enriched and subjected to further rounds of selection, creating a feedback loop that continuously improves specificity and reduces unspecific binding through iterative optimization.
2Productivity
If SELEX method is used for aptamer selection, then binding aptamers can be identified, but yield is limited
Solution Approach 1:
The patent performs preliminary bioactivity screening on aptamer candidates before final selection. By assessing bioactivity early in the process, the method identifies high-yield aptamers with desired functional properties, avoiding wasted effort on aptamers that bind but lack the required bioactivity.
Solution Approach 2:
The patent changes selection parameters from binding affinity alone to include bioactivity metrics. This parameter change enables identification of aptamers that meet both binding and functional requirements, increasing the yield of truly effective aptamers while maintaining precise bioactivity characterization.
3Reliability
If only binding affinity is used as selection criterion, then selection process is simple, but false positives increase
Solution Approach 1:
The patent merges binding affinity selection with bioactivity determination in an integrated selection process. By combining these two criteria, the method eliminates false positives that would pass binding-only selection, improving accuracy while the modular design keeps process complexity manageable.
Solution Approach 2:
The patent replaces simple mechanical binding-based selection with a more sophisticated system that incorporates biochemical activity assessment. This substitution uses enzymatic or cellular readouts to detect true functional aptamers, significantly reducing false positives despite increased process complexity.
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The invention relates to a specific SELEX based method named AGE-SELEX which includes various specific steps including at least one bioactivity determination step allowing to obtain aptamers with high bioactivity modulation properties.