Rare Molecule Detection via Affinity-MS Labeling
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Solution Overview
Problem
Current methods for detecting rare molecules in blood samples are limited by the sensitivity of conventional affinity assays and nucleic acid assays, which struggle to achieve detection below picomolar levels, and mass spectrometry techniques face issues with sample interference, sensitivity loss, and inability to work with small sample volumes, leading to inaccurate results.
Innovation Solution
The method involves enhancing the concentration of rare molecules using affinity agents with specific binding partners, followed by mass spectrometry label precursors and alteration agents to form MS labels, which are then analyzed using a porous matrix to separate and detect rare molecules and cells, allowing for high sensitivity and automation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional affinity assays are used to detect rare molecules, then the method is simple to operate, but the detection limit cannot reach sub-picomolar levels due to antibody affinity binding constant limitations
Solution Approach 1:
The patent introduces MS label precursors as intermediary molecules that bridge the affinity binding step and mass spectrometry detection. These precursors are converted to MS labels through alteration agents, enabling detection of rare molecules at sub-picomolar levels while maintaining operational simplicity through the use of standard affinity reagents like antibodies
Solution Approach 2:
The patent replaces the traditional optical or electrochemical detection systems with mass spectrometry-based detection. This substitution enables significantly improved detection limits by leveraging the high sensitivity and specificity of MS technology, which can detect molecules at femtomolar to attomolar concentrations
2Measurement precision
If nucleic acid assays with amplification are used to detect rare molecules, then detection sensitivity can be improved, but the analysis time increases to several days due to lengthy purification and amplification steps
Solution Approach 1:
The patent extracts and detects endogenous MS labels directly from rare molecules without requiring nucleic acid amplification or lengthy purification steps. By using specific affinity agents to enrich rare molecules followed by MS detection of their intrinsic labels, the method achieves high sensitivity in a much shorter time frame
Solution Approach 2:
The patent uses MS label precursors that are converted to detectable MS labels, creating a detectable copy or signal from the rare molecule itself. This approach avoids the need for multiple amplification cycles while maintaining detection sensitivity
3Measurement precision
If mass spectrometry is used to detect rare molecules, then detection sensitivity can be improved, but sample interference and background noise reduce measurement accuracy
Solution Approach 1:
The patent converts potential interference from complex biological samples into a benefit by using specific affinity agents that selectively bind to rare molecules, enriching them from the background. The subsequent MS detection of unique labels on these enriched molecules allows discrimination from background noise and interference
Solution Approach 2:
The patent uses MS labels that provide unique mass spectral signatures or 'colors' for different rare molecules. These distinct labels enable clear differentiation between target molecules and background interference, allowing accurate detection even in complex samples
4Quantity of substance
If conventional cell filtration methods are used to separate rare cells, then separation can be achieved, but cell loss occurs and contamination with non-rare cells increases due to diameter overlap between cell populations
Solution Approach 1:
The patent applies local quality by using specific affinity agents that bind to unique markers on rare cell surfaces. This targeted binding allows selective enrichment of rare cells based on their specific molecular characteristics rather than general physical properties like size, thereby avoiding loss of small rare cells and contamination from larger non-rare cells
Solution Approach 2:
The patent changes the separation parameter from physical size (diameter) to molecular specificity (marker expression). By using affinity-based recognition of cell-surface markers, the method achieves high separation efficiency and purity without the cell loss and contamination issues inherent in size-based filtration methods
5Measurement precision
If scanning microscopy is used to analyze rare cells at single cell level, then detection sensitivity can reach attomolar levels, but the analysis time exceeds 24 hours and requires manual visual examination
Solution Approach 1:
The patent replaces manual visual examination and time-consuming scanning microscopy with automated mass spectrometry-based detection. The MS system automatically detects and quantifies MS labels from rare cells, achieving attomolar detection sensitivity without requiring hours of manual analysis or specialized microscopy equipment
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 detection of rare molecules and cells at sub-attomolar levels with high sensitivity and efficiency, reducing background interference and enabling clinically accurate measurements without the need for complex preprocessing or trypsin digestion.
Implementation Method 1
an affinity agent that comprises a specific binding partner that is specific for and binds to a target rare molecule
Implementation Method 2
A retentate and a filtrate are formed by contacting the incubated sample with a porous matrix
Data Source
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AI summary
A concentrated sample having enhanced concentration of the one or more different populations of target rare molecules is incubated with, for each different population of target rare molecules, a particulate or non-particulate affinity agent that comprises a specific binding partner that is specific for and binds to a target rare molecule. The affinity agent comprises a mass spectrometry (MS) label precursor or a first alteration agent, which either facilitates the formation of an MS label from the MS label precursor or releases an entity that comprises the MS label precursor from the affinity agent. The MS label corresponds to one of the populations of target rare molecules. A second alteration agent is employed if the first alteration agent does not facilitate the formation of an MS label from the MS label precursor. MS analysis is used to determine each different MS label.