Aqueous Two-Phase Purification for Urogenital Cancer Testing
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Solution Overview
Problem
Existing methods for concentrating and purifying target analytes from clinical biological samples, such as nucleic acids, are inefficient, leading to low yield and diagnostic insensitivity due to low concentrations and excessive reagent use.
Innovation Solution
A method utilizing Aqueous Two-Phase Systems (ATPS) with polymers, salts, and surfactants to partition and concentrate target analytes, followed by solid phase media for purification, reducing reagent use and enhancing recovery efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional extraction methods are used to concentrate and isolate target analytes, then the process is simple, but the yield is low and diagnostic sensitivity is insufficient
Solution Approach 1:
The extraction process is divided into distinct functional stages: ATPS-based concentration/purification stage followed by solid-phase extraction stage. This segmentation allows each stage to specialize in specific functions, improving overall yield while maintaining workflow manageability through modular design.
Solution Approach 2:
ATPS serves as an intermediary system between the clinical sample and the solid-phase extraction medium. It performs preliminary concentration and purification, removing interfering substances that would otherwise inhibit binding, thereby improving yield without requiring complex direct binding conditions.
2Quantity of substance
If conventional extraction methods are used, then the workflow is straightforward, but the amount of binding buffer required is extremely large and unreasonable
Solution Approach 1:
ATPS performs preliminary concentration and purification of the target analyte before it reaches the solid-phase extraction stage. This preliminary action reduces the volume of sample that requires binding buffer, making the overall reagent consumption reasonable and operationally feasible.
Solution Approach 2:
The method changes the concentration parameter of the target analyte using ATPS before solid-phase extraction. By concentrating the sample in the ATPS phase, the subsequent binding step requires proportionally less binding buffer, making the workflow practical.
3Measurement precision
If target analytes are present at extremely low concentrations, then diagnostic sensitivity can be achieved, but the yield of purified analyte becomes insufficient for analysis
Solution Approach 1:
ATPS selectively extracts and concentrates the target analyte from the complex clinical sample matrix, separating it from interfering substances. This extraction step achieves both concentration (improving yield) and purification (maintaining sensitivity) simultaneously.
Solution Approach 2:
The ATPS creates a localized high-concentration phase where the target analyte accumulates. This local concentration effect allows sufficient yield to be obtained while maintaining the purity needed for sensitive diagnostic analysis.
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 concentrates and purifies target analytes, minimizing interference from proteins and salts, and allows for efficient downstream analysis with reduced reagent use, suitable for diagnostic applications like bladder cancer and HPV detection.
Implementation Method 1
combining the clinical biological sample with a first aqueous two-phase system (ATPS) composition including a polymer, a salt component including at least one salt, a surfactant, or any combination thereof dissolved in an aqueous solution to form a target-rich phase solution and a target-poor phase solution
Implementation Method 2
contacting the target rich phase from step (b), the second target-rich phase from step (c) or the mixed solution from step (d) with a solid phase medium configured to selectively bind the target analyte such that the solid phase medium binds to the target analyte
Data Source
AI summary
The present disclosure relates to methods, compositions, and kits for concentrating and purifying at least one target analyte from a clinical biological sample. In some embodiments, the methods involve one or more aqueous two-phase system (ATPS) compositions and at least one solid phase medium. Some embodiments provide a kit comprising one or more ATPS compositions, a binding buffer; and a solid phase medium. Other embodiments provide methods of treating cancers or infectious diseases in a patient in need thereof.