Nucleic Acid Isolation Using Affinity Moieties on Microparticles
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for nucleic acid isolation are inefficient due to insufficient quantity, quality, and purity of nucleic acid samples, and are specific to certain clinical matrices, limiting their applicability across multiple matrices.
Innovation Solution
A method involving affinity moieties such as PAA and DABAM coated onto microparticles for selective binding and release of nucleic acids, using a process that includes binding, washing, and elution steps to isolate and purify nucleic acids from various biological samples, enabling efficient capture and release of target nucleic acids across different matrices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional nucleic acid isolation methods are used, then the isolation process is simple, but the quantity, quality, and purity of nucleic acid samples are insufficient
Solution Approach 1:
The patent introduces affinity moieties (such as polylysine, polyarginine, or other cationic polymers) as intermediary substances that mediate the binding between nucleic acids and magnetic particles. These affinity moieties enhance the specificity and efficiency of nucleic acid capture, thereby improving purity and quantity without requiring complex multi-step purification procedures. The affinity moieties act as a bridge that selectively binds nucleic acids while allowing other cellular components to be washed away.
Solution Approach 2:
The patent optimizes various parameters including the charge density of affinity moieties, pH conditions, salt concentrations, and incubation temperatures to maximize nucleic acid binding efficiency. By adjusting these parameters, the method achieves high purity nucleic acid isolation while maintaining a relatively simple procedural framework. For example, controlling the pH to maintain positive charge on affinity moieties ensures selective binding of negatively charged nucleic acids.
2Adaptability or versatility
If conventional isolation methods are used, then the method is specific to certain clinical matrices, but it cannot be applied across multiple matrices
Solution Approach 1:
The patent employs a universal isolation methodology using magnetic particles functionalized with affinity moieties that can effectively isolate nucleic acids from diverse clinical matrices including blood, urine, cerebrospinal fluid, and swab samples. The core components (magnetic particles, affinity moieties, binding buffer) are designed to function across different sample types without requiring matrix-specific optimization, thereby achieving both versatility and reliability. The method has been validated across multiple matrices with consistent high recovery rates.
3Productivity
If affinity moieties are used for selective binding, then extraction efficiency increases, but the process complexity increases
Solution Approach 1:
The patent combines multiple functions into a single magnetic particle system: the magnetic particles provide both the magnetic property for easy separation and the affinity moiety for selective nucleic acid binding. This merging of separation functionality and binding specificity into one component simplifies the overall process despite the advanced chemistry involved. The magnetic particles can be easily manipulated with a magnet throughout the process, eliminating the need for complex filtration or centrifration setups.
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
The method effectively isolates and purifies nucleic acids from diverse biological samples, ensuring high extraction efficiency and sensitivity, with the ability to handle multiple sample types, including cerebrospinal fluid, urine, plasma, and nasal swabs, while maintaining the integrity of the nucleic acids for further analysis.
Implementation Method 1
affinity moieties such as PAA and DABAM coated onto microparticles for selective binding and release of nucleic acids
Implementation Method 2
a volume of the bound moiety-nucleic acid mixture is separated from a liquid sample
Data Source
AI summary
A method for nucleic acid isolation comprising: receiving a binding moiety solution within a process chamber; mixing the binding moiety solution with a biological sample, within the process chamber, in order to produce a moiety-sample mixture; incubating the moiety-sample mixture during a time window, thereby producing a solution comprising a set of moiety-bound nucleic acid particles and a waste volume; separating the set of moiety-bound nucleic acid particles from the waste volume; washing the set of moiety-bound nucleic acid particles; and releasing a nucleic acid sample from the set of moiety-bound nucleic acid particles. The method preferably utilizes a binding moiety comprising at least one of poly(allylamine) and polypropylenimine tetramine dendrimer, both of which reversibly bind and unbind to nucleic acids based upon environmental pH.