Micro-channel Cell Capture with Cross-linked Binding Partners
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Solution Overview
Problem
Current methods for isolating and analyzing circulating tumor cells from biological samples face challenges such as non-specific binding, harsh analysis conditions that degrade binding partners, and limitations of cytokeratin markers for identifying malignancy.
Innovation Solution
A micro-channel device and method that pre-labels samples with a binding partner specific to the target cell, using a combination of binding entities to enhance capture and analysis, including the use of antibody cocktails and cross-linking reagents to stabilize cells during analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If washing steps are performed to remove non-specific binding, then non-specific binding is minimized, but the binding partner may degrade, lose conformational structure, or become detached
Solution Approach 1:
The patent introduces cross-linking reagents as intermediary substances that form covalent bonds between the binding partner and the solid support, creating a stable linkage that prevents detachment during washing steps while maintaining specific binding capability
Solution Approach 2:
The patent applies cross-linking treatment before performing washing steps, stabilizing the binding partner in advance to prevent degradation and detachment that would otherwise occur during subsequent washing procedures
2Productivity
If harsh staining and hybridization procedures are used for in situ analysis, then analysis can be performed, but the binding partner may degrade or lose conformational structure
Solution Approach 1:
The patent stabilizes the binding partner through cross-linking before subjecting it to harsh staining and hybridization conditions, ensuring structural integrity is maintained throughout the analysis process
Solution Approach 2:
The patent provides protective stabilization to the binding partner in advance of harsh analysis procedures, cushioning against potential damage from denaturing conditions during staining and hybridization
3Measurement precision
If cytokeratin staining is used to identify circulating tumor cells, then malignancy can be evaluated, but cells without cytokeratin expression cannot be detected
Solution Approach 1:
The patent employs binding partners that can recognize multiple different cell surface markers beyond just cytokeratin, enabling the system to detect and analyze various types of circulating tumor cells with different phenotypic characteristics through a single platform
Solution Approach 2:
The patent changes the detection parameter from relying solely on cytokeratin expression to using alternative cell surface markers, thereby expanding the detection capability to include malignancy types that do not express cytokeratin
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
Improves the capture efficiency of rare cells like circulating tumor cells and allows for effective analysis of malignancy without relying on cytokeratin expression, providing a more stable and efficient method for post-capture analysis.
Implementation Method 1
The loading mixture may comprise a biological sample suspected of containing a target, such as a target cell, and also comprises a first binding entity. The first binding entity specifically binds to the target (e.g., a target entity on a target cell). The surface of the micro-channel is coated with a second binding entity that specifically binds, directly or indirectly, to the first binding entity.
Data Source
AI summary
The present invention provides a device for isolating target biomolecules or cells from samples, particularly biological samples. In particular, the device comprises a loading mixture, which contains the biological sample and a first binding entity that specifically binds to the target biomolecule or target cell; and a micro-channel coated with a second binding entity that binds directly or indirectly to the first binding entity. Methods of capturing, detecting, and/or evaluating target biomolecules or target cells (e.g. cancer cells) in biological samples are also disclosed.