Urine Cell-Free DNA Isolation with ATPS and Solid-Phase Binding
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional methods for concentrating and purifying target analytes from clinical biological samples, such as nucleic acids, are inefficient, leading to low yields and insufficient diagnostic sensitivity due to low concentrations and excessive reagent use.
Innovation Solution
A method utilizing Aqueous Two-Phase Systems (ATPS) and solid phase media, involving multiple steps of partitioning and binding to concentrate and purify target analytes, minimizing reagent use and interference from salts and proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional extraction methods are used, then the process is simple, but the yield of purified target analytes is low and diagnostic sensitivity is insufficient
Solution Approach 1:
The extraction process is divided into distinct functional stages: ATPS partitioning step to separate target analytes from bulk matrix, solid phase binding step to concentrate and purify, and elution step to recover purified analytes. This segmentation allows each step to be optimized independently, improving overall yield while maintaining workflow manageability.
Solution Approach 2:
Aqueous two-phase system (ATPS) acts as an intermediary between the clinical biological sample and the solid phase medium. The ATPS selectively partitions target analytes into a target-rich phase, removing interfering salts and proteins before binding to the solid phase, thereby enabling higher yield and purity without complicating the overall workflow.
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:
The ATPS partitioning step is performed as a preliminary action before the binding step. By pre-concentrating and pre-purifying the target analytes in the ATPS target-rich phase, the subsequent binding step requires significantly reduced amounts of binding buffer, eliminating the need for excessive buffer volumes while maintaining extraction efficiency.
3Measurement precision
If conventional extraction methods are used, then reagent use is high, but the diagnostic sensitivity and specificity are insufficient
Solution Approach 1:
The ATPS system selectively extracts and removes interfering substances (salts, proteins) from the sample matrix before the binding step. This extraction of interferents concentrates the target analytes and eliminates substances that would otherwise compete for binding sites or interfere with detection, thereby improving diagnostic sensitivity and specificity while reducing the need for excessive reagents.
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 achieves higher recovery efficiency and concentration of target analytes like cell-free DNA, enabling effective diagnostic tests for cancers and infectious diseases with reduced reagent use and equipment requirements.
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.