Amide Stabilizing Composition for Multimodal Liquid Biopsy
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Solution Overview
Problem
Current liquid biopsy methods face challenges in the reliable stabilization and analysis of multiple biological targets from a single cell-containing bodily fluid sample, particularly due to the rarity of cells like CTCs and the short half-life of CTCs in circulation, as well as the interference of formaldehyde-based stabilization methods with nucleic acid isolation.
Innovation Solution
A method involving a stabilizing composition containing primary, secondary, or tertiary amides, poly(oxyethylene) polymers, and apoptosis inhibitors is used to stabilize cell-containing bodily fluids, allowing for the simultaneous enrichment and analysis of rare cells, extracellular nucleic acids, and extracellular vesicles, enabling the preservation of biological targets for extended periods without cross-linking issues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If formaldehyde-based stabilization methods are used, then cell preservation is improved, but nucleic acid isolation is interfered with
Solution Approach 1:
The patent changes the chemical parameters of the stabilization composition by replacing formaldehyde with a combination of amide compounds (such as succinimide, N-acetyl-L-cysteine) and apoptosis inhibitors. This parameter change maintains cell preservation effectiveness while eliminating the harmful cross-linking effect that interferes with nucleic acid isolation, thus resolving the contradiction between reliable cell preservation and ease of nucleic acid isolation.
2Loss of information
If multiple biological targets are analyzed from a single sample, then diagnostic information is improved, but analysis complexity is increased
Solution Approach 1:
The patent applies segmentation by dividing the analysis process into distinct modules: cell enrichment module, nucleic acid extraction module, and vesicle isolation module. Each module can process different biological targets independently from the stabilized sample, allowing comprehensive diagnostic information to be obtained while managing complexity through modular, standardized procedures rather than a single complex analysis pipeline.
Solution Approach 2:
The stabilization composition serves multiple functions simultaneously: it preserves cell morphology, maintains nucleic acid integrity, prevents apoptosis, and stabilizes extracellular vesicles. This multi-functionality allows a single sample stabilization step to support multiple downstream analyses (CTC detection, ctDNA extraction, EV isolation), reducing overall analysis complexity while maximizing diagnostic information retrieval.
3Measurement precision
If CTC enrichment is performed, then detection sensitivity is improved, but cell integrity is compromised
Solution Approach 1:
The patent applies preliminary action by performing stabilization treatment on the blood sample immediately upon collection, before any enrichment or manipulation steps. This pre-stabilization creates a protective environment that maintains cell integrity throughout subsequent CTC enrichment procedures, allowing detection sensitivity to be improved through enrichment while the previously established stabilization prevents cell degradation or artifact formation.
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 approach enables the reliable and efficient enrichment and analysis of multiple biological targets from a single sample, improving the sensitivity and robustness of liquid biopsy analyses by maintaining the integrity of nucleic acids and cells, even after prolonged storage, and allowing for the detection of tumor cells with high sensitivity.
Implementation Method 1
contacting a cell-containing bodily fluid with a stabilizing composition comprising one or more of the following stabilizing agents: (a) at least one primary, secondary or tertiary amide, (b) at least one poly(oxyethylene) polymer, and/or (c) at least one apoptosis inhibitor
Implementation Method 2
allowing for the simultaneous enrichment and analysis of rare cells, extracellular nucleic acids, and extracellular vesicles, enabling the preservation of biological targets for extended periods without cross-linking issues
Data Source
AI summary
A method for stabilizing and isolating multiple biological targets comprised in a cell-containing bodily fluid, said method comprising (A) contacting a cell-containing bodily fluid with a stabilizing composition comprising one or more of the following stabilizing agents: (a) at least one primary, secondary or tertiary amide, (b) at least one poly(oxyethylene) polymer, and/or (c) at least one apoptosis inhibitor, thereby providing a stabilized cell-containing bodily fluid sample; (B) keeping the stabilized cell-containing bodily fluid sample for a stabilization period; and (C) processing the stabilized cell-containing bodily fluid sample in order to enrich three or more biological targets selected from the group consisting of —a cell subpopulation, —extracellular nucleic acids, —extracellular vesicles and —intracellular nucleic acids from the stabilized cell-containing bodily fluid. The method is advantageous and enables the multimodal analyses of different biological targets from a single stabilized cell-containing body fluid sample.