Multiplex Aptamer Selection for Vitamin C and HRP2 Binding
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Solution Overview
Problem
Existing methods for generating functional biomolecules, such as aptamers, are inefficient and costly when targeting multiple molecules simultaneously, and there is a need for high-capacity, multiplexed identification and diagnostic applications for molecules like vitamin C and malaria antigens.
Innovation Solution
A method for generating functional nucleic acids and peptides that bind to multiple targets simultaneously using a single reaction volume, involving a library of biomolecules applied to an array with targets, followed by partitioning, tagging, and amplification to identify specific binders, utilizing SELEX and phage display techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If separate identification processes are used for each target molecule, then specificity and binding affinity can be optimized for each individual target, but the time, cost, and complexity increase significantly
Solution Approach 1:
The patent combines multiple target identification processes into a single multiplexed assay. Multiple target molecules are simultaneously presented to the aptamer library in one reaction volume, allowing parallel selection of aptamers binding to different targets (vitamin C, HRP2, and other molecules) without requiring separate experiments for each target.
Solution Approach 2:
The invention creates a universal selection system that can identify aptamers for multiple different targets using the same experimental platform. The method uses a common library of randomized nucleic acids and standardized selection procedures that work across diverse target types, eliminating the need for target-specific optimization of the entire selection process.
2Reliability
If multiple separate experiments are conducted to generate aptamers for different targets, then each aptamer can be optimized for its specific target, but the cost and resource requirements increase
Solution Approach 1:
The patent merges multiple aptamer generation experiments into one cost-effective process. By presenting multiple targets simultaneously to a single aptamer library and using combined amplification and selection steps, the method reduces reagent consumption, labor costs, and facility usage compared to running separate experiments for each target.
Solution Approach 2:
The method recovers and reuses components across multiple selection rounds and targets. The aptamer library is amplified and reused for subsequent selection cycles, and non-binding aptamers are discarded while binding aptamers are recovered and further selected, maximizing the utility of each experimental input.
3Measurement precision
If traditional SELEX methods are used for each target separately, then detailed characterization can be performed for each aptamer-target pair, but the overall process becomes excessively complex and time-consuming
Solution Approach 1:
The patent segments the complex multiplexed selection process into distinct, manageable stages: (1) simultaneous presentation of multiple targets to the aptamer library, (2) washing to remove non-binding aptamers, (3) combined amplification of bound aptamers, and (4) iterative selection rounds. This segmentation reduces operational complexity while maintaining the ability to characterize multiple aptamer-target interactions.
Solution Approach 2:
The method uses nucleic acid amplification to generate copies of binding aptamers for further selection and characterization. This copying mechanism allows the same amplification and selection protocol to be applied across multiple targets simultaneously, reducing the need for separate detailed characterization experiments for each target-aptamer pair.
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 efficient, cost-effective, and time-saving generation of functional ligands for multiple targets, facilitating diagnostics and therapeutic applications by reducing the need for separate target identification processes.
Implementation Method 1
Aptamers are typically characterized by binding to their target molecules via non-Watson-Crick (i.e. non-hybridization) mechanisms, such as by intermolecular forces resulting from the secondary or tertiary structure of the aptamer
Implementation Method 2
Aptamers are commonly identified by an in vitro method of selection sometimes referred to as Systematic Evolution of Ligands by EXponential enrichment or 'SELEX'
Implementation Method 3
utilizing SELEX and phage display techniques
Data Source
AI summary
The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules, such as vitamin C or malaria histidine-rich protein II (HRP2).