Pre-mixed Reagent Vessel for Rapid Proteomic Sample Digestion
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Solution Overview
Problem
Current proteomic sample preparation methods for mass spectrometry are time-consuming, require significant user involvement, and lack reproducibility due to sequential steps and multiple reagent additions, which hinder high-throughput analysis.
Innovation Solution
A method involving a closeable reaction vessel with pre-measured reagents including immobilized proteolytic enzymes, reducing agents, and alkylating agents, where the sample is concurrently reacted with these agents by heating, sonicating, or applying pressure, significantly reducing processing time to between 5 minutes and 90 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional sequential proteomic sample preparation methods are used, then comprehensive protein characterization is achieved, but processing time exceeds 16 hours and user involvement is significant
Solution Approach 1:
The patent combines multiple sequential steps (cell lysis, protein reduction, alkylation, and digestion) into a single concurrent reaction step. The reagent mixture contains all necessary components that act simultaneously on the protein sample, reducing processing time from 16+ hours to 5-90 minutes while maintaining comprehensive protein characterization capability.
Solution Approach 2:
The reagents are pre-measured and pre-mixed in the reaction vessel before the sample is introduced. This preliminary preparation eliminates the need for multiple sequential reagent additions and user involvement during the reaction process, enabling automated high-throughput processing while ensuring reproducible results.
2Reliability
If multiple reagent additions are performed sequentially, then complete protein modification is achieved, but user involvement increases and reproducibility decreases
Solution Approach 1:
Multiple reagents (proteolytic enzyme, reducing agent, alkylating agent) are combined into a single pre-measured reagent mixture that is added once to the sample. This eliminates multiple user intervention steps, improves reproducibility by eliminating variability in sequential additions, and simplifies operation while maintaining complete protein modification.
Solution Approach 2:
The pre-measured reagent mixture is designed to automatically perform all necessary protein modifications when activated by a single input (such as sample introduction or mild heating). The system self-regulates the reaction progression without requiring user monitoring or intervention, enhancing both ease of operation and reproducibility.
3Loss of time
If rapid digestion kits with multiple reagent additions are used, then processing time is reduced to 1-3.5 hours, but user involvement remains significant
Solution Approach 1:
The patent merges all necessary reagents into a single pre-measured mixture that enables concurrent protein modification and digestion. This single addition approach reduces processing time to 5-90 minutes (faster than 1-3.5 hours) while eliminating the need for multiple user additions, achieving both time reduction and automation.
Solution Approach 2:
The invention changes the parameter of reagent delivery from multiple sequential additions to a single concurrent addition of a pre-mixed reagent solution. This parameter change enables the reaction to proceed faster (5-90 minutes) while requiring minimal user involvement, as the pre-measured mixture is prepared in advance and added as one step.
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 enhances digestion efficiency and alkylation efficiency, achieving results comparable to traditional methods while significantly reducing sample preparation time and user intervention, maintaining high reproducibility and stability across varying protein amounts and storage conditions.
Implementation Method 1
an immobilized proteolytic enzyme
Implementation Method 2
proteolytic digestion of a protein-containing sample
Implementation Method 3
a reducing agent
Implementation Method 4
an alkylating agent
Implementation Method 5
activating the contents of the reaction vessel
Implementation Method 6
sonicating the sample
Data Source
AI summary
A method for preparing a protein-containing sample for analysis by mass spectrometry includes introducing the sample into a reaction vessel. The reaction vessel contains a reagent mixture including pre-measured quantities of an immobilized proteolytic enzyme, a reducing agent and an alkylating agent. The contents of the reaction vessel are activated by heating or by sonication.