Integrated Microfluidic Chip for Protein Purification and Cryo-EM Deposition
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Solution Overview
Problem
Current laboratory techniques for protein purification and cryogenic electron microscopy (cryo-EM) preparation are time- and labor-intensive, requiring multiple meticulous interventions and are not optimized for low copy eukaryotic proteins, with existing microfluidic devices often lacking integrated deposition capabilities on a single chip.
Innovation Solution
A stand-alone microfluidic lab-on-a-chip system for protein purification, integrating a purification device, detection system, and controlled deposition onto a microscopy grid, allowing minimal operator intervention and reduced procedural steps, particularly suitable for cryo-EM applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If standard laboratory techniques for protein purification and cryo-EM preparation are used, then purification can be achieved, but the process is time- and labor-intensive requiring multiple meticulous interventions
Solution Approach 1:
The patent combines multiple purification steps (affinity purification, size exclusion chromatography) and the deposition step into a single integrated microfluidic chip device. This merging of functions into one automated system eliminates the need for multiple separate manual operations, directly reducing preparation time and increasing throughput by allowing continuous processing without operator intervention between steps.
Solution Approach 2:
The microfluidic chip is pre-configured with integrated purification modules and deposition mechanisms before sample introduction. The system performs preliminary setup of all necessary purification steps and automation controls in advance, enabling the sample to flow through the entire purification and deposition process automatically without requiring real-time manual adjustments or interventions during processing.
2Ease of operation
If existing microfluidic devices are used, then miniaturization is achieved, but they lack integrated deposition capabilities requiring operator intervention
Solution Approach 1:
The microfluidic chip is designed as a universal platform that performs multiple functions: affinity purification, size exclusion chromatography, and direct deposition onto microscopy grids. This multi-functional integration allows a single device to replace multiple separate instruments, achieving high automation while the modular design manages complexity by combining standardized purification modules with a simple deposition mechanism.
3Quantity of substance
If bulk purification columns are used, then purification capacity is sufficient, but starting volumes must be in the order of few milliliters which is not suitable for low copy proteins
Solution Approach 1:
The patent implements nested purification modules within the microfluidic chip, where affinity purification beads are contained within micro-chambers that are themselves integrated into the larger chip structure. This nested arrangement allows sufficient purification capacity to be achieved in a miniaturized format, enabling effective purification of low copy proteins from very small starting volumes (nanoliters to microliters) while maintaining reliability through the concentrated efficiency of the nested design.
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 system significantly reduces preparation time and simplifies the process, enabling efficient purification and deposition of proteins/protein complexes directly onto a microscopy grid, facilitating high-throughput cryo-EM analysis with minimal sample volumes.
Implementation Method 1
The channel system (25) internally comprises a first purification device (14) for purifying one or more proteins or protein complexes of interest from a sample
Implementation Method 2
The first purification device (14) comprises at least one inlet (15) for introducing or removing purification agents
Implementation Method 3
The channel system (25) comprises a first detection section (16) suitable for detection of a label or marker in the sample, preferably wherein the first detection section (16) is suitable for fluorescence detection
Implementation Method 4
The channel system (25) internally comprises a first on-chip valve system downstream from the first purification device (14), and preferably downstream from the first detection section (16), towards the chip outlet (22)
Implementation Method 5
The chip outlet (22) is provided with a deposition means suitable for cryo-EM grid sample deposition
Data Source
AI summary
The current invention relates to an improved microfluidic device for protein/protein complex purification, as well as an associated methodology and system.In particular it relates to systems and methods for electron microscopy, preferably cryogenic electron microscopy (cryo-EM), on protein samples, comprising: a. at least one microfluidic chip with inlet and outlet and purification device; b. at least one illumination means and detection means for, preferably fluorescence, measurements on the microfluidic chip; c. a pumping system adapted for operable connection to the microfluidic chip, and configured for controlling flow in the operably connected microfluidic chip; d. a microscopy grid, preferably a cryogenic electron microscopy (cryo-EM) grid, for holding fluid samples; e. preferably a cryogenic container, for a cryogenic coolant; f. preferably a transport system for moving the cryo-EM grid between a position for receiving a fluid sample from the microfluidic chip and the cryogenic container; g. a control system, preferably a processor, which receives information on the measurements from the detection means, configured for controlling the pumping system at least based on said information, and further configured for controlling the illumination and detection means and preferably also for controlling the transport system.