Thiamine-Antithiamine Releasable Conjugate for Cell Separation
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Solution Overview
Problem
Existing cell detection and separation techniques face challenges with residual labeling due to the release of biotin-based conjugates, which can cause cross-talk and interfere with downstream applications, necessitating a more reliable and orthogonal tag/anti-tag system.
Innovation Solution
A conjugate system utilizing thiamine and anti-thiamine as binding partners, allowing for specific labeling and reversible binding, enabling efficient detection and release of target moieties without cross-reactions, using a non-covalent bond that can be disrupted with a release agent, facilitating further labeling and detection strategies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If biotin/anti-biotin conjugate system is used for labeling, then specific binding and detection are achieved, but cross-talk and residual labeling occur causing interference with downstream applications
Solution Approach 1:
The patent extracts the problematic biotin component from the tag/anti-tag system and replaces it with thiamine. This removes the source of cross-talk and residual labeling issues while preserving the specific binding functionality needed for detection.
Solution Approach 2:
The patent changes the chemical parameter of the binding system from biotin-based to thiamine-based. This parameter change results in a new binding system with different properties that does not exhibit cross-talk or residual labeling, thereby resolving the harmful effects while maintaining specificity.
2Stability of the object's composition
If irreversible covalent bonding is used for labeling, then stable detection is achieved, but release of label for downstream applications becomes difficult
Solution Approach 1:
The patent transitions from a static irreversible covalent bond to a dynamic reversible non-covalent bond. This allows the label to be stable during detection but easily released when needed for downstream applications, providing operational flexibility.
Solution Approach 2:
The patent changes the bonding parameter from covalent (irreversible) to non-covalent (reversible). This parameter change enables the label to remain stable during detection while allowing controlled release for subsequent labeling strategies, resolving the contradiction between stability and ease of release.
3Device complexity
If single labeling strategy is used, then simplicity is maintained, but ability to perform sequential sorting and molecular diagnostic is limited
Solution Approach 1:
The patent segments the labeling process into distinct phases: initial labeling with thiamine/anti-thiamine conjugate for detection, followed by potential release and re-labeling steps. This segmentation enables sequential sorting strategies and molecular diagnostics while maintaining protocol simplicity through modular design.
Solution Approach 2:
The reversible nature of the non-covalent bond creates dynamic labeling capability, allowing the system to transition between bound and unbound states. This dynamic behavior enables multiple labeling strategies and sequential applications without increasing overall protocol complexity.
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 thiamine/anti-thiamine system effectively allows for the detection and isolation of target cells with high efficiency, achieving greater than 80% label removal, reducing interference and enabling multiple labeling and detection strategies without cross-talk, thus enhancing the accuracy and reliability of downstream applications.
Implementation Method 1
B and C are non-covalently bound to each other
Data Source
AI summary
The invention is directed to a conjugate complex for detecting a target moiety in a sample of biological specimens having the general formula (I) An - Bm...Cq-Xo (I) with A: antigen recognizing moiety; B: first binding moiety C second binding moiety X: detection moiety; n, m, q, o integers between 1 and 100, wherein B and C are non-covalently bound to each other characterised in that B comprises a thiamine unit and C is a moiety recognizing thiamine. Futher, the invention is directed to a method detecting a target moiety in a sample of biological specimens with a conjugate complex having the general formula (I).