Microfluidic Single-Cell Glycan Profiling via Lectin Binding
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for single-cell glycan profiling are limited by high costs, low throughput, and inability to analyze individual living cells, as well as a lack of tools for unambiguous determination of glycan structures and modifications, particularly in clinical and biological samples.
Innovation Solution
A method using carbohydrate-binding molecules like lectins, antibodies, and aptamers applied in a microfluidic platform to detect glycan binding and reconstruct glycan profiles at the single-cell level through binding intensity measurement and machine learning algorithms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional glycan analysis technologies (FACS, MS, HPLC) are used for bulk cell populations, then glycan composition can be identified, but the methods are costly, tedious and time-consuming with low throughput
Solution Approach 1:
The invention segments the glycan analysis process into microfluidic droplets, with each droplet containing a single cell and undergoing independent glycan extraction and analysis. This segmentation enables parallel processing of thousands of individual cells simultaneously, transforming low-throughput sequential analysis into high-throughput parallel analysis while maintaining single-cell resolution
Solution Approach 2:
The invention introduces an integrated microfluidic platform as an intermediary system that automates and integrates multiple manual steps (cell lysis, glycan extraction, chromatography) into a single automated workflow. This intermediary platform eliminates the need for tedious manual operations and enables high-throughput processing while preserving measurement precision
2Measurement precision
If existing single-cell glycan profiling methods are used, then individual cell analysis is possible, but the methods are not appropriate for profiling single-cell surface glycome and cannot handle multiple sequential probing
Solution Approach 1:
The microfluidic platform is designed with universal functionality to perform multiple sequential operations on the same single-cell lysate: glycan extraction, chromatographic separation, and detection. The system can sequentially probe different glycan epitopes using multiple lectins in a single integrated workflow, enabling comprehensive single-cell glycomics analysis that previous methods could not achieve
3Measurement precision
If glycan analysis technologies are used for large cell populations, then composition identification is possible, but the cells are destroyed and unable to handle multiple sequential probing
Solution Approach 1:
The invention segments the cell population into individual single-cell lysates within separate microfluidic droplets. Each droplet maintains its own glycan extract that can undergo multiple sequential lectin probing operations without affecting other cells. This segmentation enables automated sequential probing of the same sample while preserving the integrity of individual cell-derived glycans
Solution Approach 2:
The microfluidic platform enables continuous automated processing where glycan extracts from single cells undergo sequential lectin binding reactions without manual intervention. The system continuously flows different lectins through the droplets, allowing multiple probing operations to occur in sequence on the same sample, thereby increasing automation extent while maintaining measurement precision
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 robust, affordable, and reliable single-cell glycan profiling, providing high-resolution structural information on glycan structures and modifications, facilitating diagnostics and therapeutic applications.
Implementation Method 1
The binding of a lectin to a glycan can be directly measured or indirectly measured through a detection tag on the lectin, the glycan, or a component of the complex formed by the lectin and the glycan.
Data Source
AI summary
A transformative method to profile the glycome in individual cells by leveraging computational biology tools with lectin or similar profiling technologies. Robust and accurate reconstruction glycomes with high-resolution glycan structure information for biological samples, including at the single cell level. Tools such as single-clone analysis andjoint-clone analysis, which may be used to assist researchers in analyzing single cell glycoprofiled samples, which identify how glycosylation variation across cells impact the cellular phenotypes. Single cell glycoprofiling using lectins is practically implemented to provide high resolution of the glycan structure information. Glycan profiling techniques having a wide range of biological applications from embryonic development to cancer and infectious disease due to high throughput, low cost, and robust reliability.