Single Cell Protein Analysis via Droplet Segmentation
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Solution Overview
Problem
Current diagnostic methods are unable to effectively monitor sub-population and cell-level variations within a sample, often providing only average measurements that mask important heterogeneities, such as those found in diseases like cancer or doping in sports.
Innovation Solution
A method involving contacting a cell with a support in a droplet, labeling biomolecules, and detecting post-translational modifications using sequencing by degradation, allowing for the isolation and measurement of specific biomolecules at the single-cell level.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sample-level average measurements are used (mass spectrometric, ELISA, immunohistological analyses), then the measurement process is simple and provides a single abundance value per target, but sub-population and cell-level variations within the sample are lost
Solution Approach 1:
The sample is segmented into individual cells, each encapsulated in a separate droplet with a magnetic bead. This segmentation enables single-cell resolution analysis while managing complexity through parallel processing of many individual cells rather than analyzing one complex sample at a time.
Solution Approach 2:
Magnetic beads serve as intermediaries that capture and concentrate polypeptides from individual cells. The beads mediate between the single cell and the detection system, enabling precise measurement while simplifying the overall process through a standardized intermediate component.
2Loss of information
If single cell analysis is performed to detect sub-population variations, then detailed cellular heterogeneity information is obtained, but the complexity of the analysis system increases
Solution Approach 1:
By segmenting the sample into individual cell-droplet-bead units, the system preserves all cellular heterogeneity information without loss. Each cell's unique polypeptide profile is captured separately, maintaining the full spectrum of biological variation present in the original sample.
Solution Approach 2:
The system changes the scale parameter from population-level averaging to single-cell resolution. This parameter change enables detection of subtle cellular heterogeneity while the use of standardized magnetic beads and droplets keeps the system complexity manageable through scalability.
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
Enables the precise measurement and identification of biomolecules at the sub-population level, providing detailed insights into cellular variations that can be crucial for personalized diagnostics and monitoring.
Implementation Method 1
detecting the at least one labeled biomolecule by sequencing by degradation
Implementation Method 2
permeabilizing the cell within the droplet, thereby bringing the polypeptide in contact with the bead
Implementation Method 3
wherein upon the polypeptide coming in contact with the bead, the polypeptide is coupled to the bead
Implementation Method 4
the first reactive moiety and the second reactive moiety form a covalent bond to form a conjugate comprising the support and the at least one biomolecule
Data Source
AI summary
Methods and compositions for partitioning and analyzing cells are provided herein. The partitioning methods include encapsulating single cells in droplets, enabling biomolecule analysis at the single cell level. Further to this concept, multiple biomarkers can be quantified from single red blood cells, including methods which may determine whether a subject has undergone autologous blood transfer.