Functional Aptamer Selection Using Cell-Response Reporter Screening
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing aptamer selection processes fail to enrich for aptamers that induce a desired cellular function, leading to inefficiencies in identifying aptamers capable of modulating target cell functions effectively.
Innovation Solution
A method involving contacting target cells with aptamer cluster particles, detecting cell-aptamer complexes undergoing a specific function, and enriching aptamers that induce desired cellular effects, such as apoptosis or immune modulation, through flow cytometry and amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional binding-based aptamer selection processes are used, then aptamers that bind to target cells can be identified, but aptamers that induce desired cellular functions are not enriched
Solution Approach 1:
The patent introduces a reporter molecule as an intermediary that mediates between the aptamer-target cell interaction and the detection system. The reporter molecule converts the biological function (e.g., apoptosis, proliferation) into a detectable signal, enabling the selection process to specifically enrich for functionally active aptamers rather than just binders
Solution Approach 2:
The patent changes the detection parameter from simple binding presence to functional outcome measurement. By monitoring cellular responses (apoptosis, proliferation, activation) rather than just binding events, the selection process shifts from enriching binders to enriching functionally active aptamers
2Ease of manufacture
If aptamers are selected based on binding alone, then binding aptamers are identified, but functional effects on target cells cannot be guaranteed
Solution Approach 1:
The patent implements a feedback mechanism where the cellular functional response is measured and used to guide the selection process. Aptamers that induce the desired functional effect are detected via the reporter signal and enriched for subsequent rounds, creating a feedback loop that continuously improves functional enrichment while maintaining process simplicity
3Measurement precision
If weakly binding aptamers are used, then specific target recognition is achieved, but functional enrichment is insufficient
Solution Approach 1:
The patent performs preliminary functional screening by incorporating the reporter system at the beginning of the selection process. This allows weakly binding aptamers with functional activity to be identified and enriched early, rather than requiring strong binders, thereby improving functional aptamer yield while maintaining target recognition specificity
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
Enriches aptamer libraries for aptamers that induce desired cellular functions, facilitating the identification of functional aptamers useful as therapeutics, with up to 1.5-fold functional enrichment.
Implementation Method 1
detecting the cell-aptamer cluster particle complexes undergoing the cell function
Implementation Method 2
detecting the cell-aptamer cluster particle complexes undergoing the cell function
Data Source
AI summary
The present disclosure describes compositions and methods for selection functional aptamers. In certain embodiments, provided herein are methods of using aptamer cluster-containing particles to identify functional aptamers from an aptamer library. In certain embodiments, provided herein are functionally enriched populations of aptamers. In certain embodiments, provided herein are methods for selecting an aptamer for use in personalized cancer treatment and methods for preparing a tumor delivery system. In certain embodiments, provide herein are compositions comprise the aptamer cluster-containing particles, target cells (e.g., cancer cells, immune cells, etc.) and/or a detectable indicator of cellular function (e.g., a fluorescent indicator of apoptosis, cell proliferation, gene or protein expression, etc.).