Immobilized Reagent Cartridge for Peptide Analysis
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Solution Overview
Problem
Conventional methods for peptide and protein analysis by LC-MS face issues with reagent residues and enzyme contamination, leading to performance deterioration and cross-contamination, especially when using reusable columns or immobilized enzyme dispersions.
Innovation Solution
A preanalysis treatment method involving immobilized reagents on supports for reducing agents and enzymes, allowing for easy separation from the sample, thereby reducing reagent mixing and facilitating disposable systems to prevent cross-contamination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional enzymatic digestion with soluble enzyme solution is used, then peptide and protein fragmentation is achieved, but reagent residues and enzyme fragments mix into the sample causing performance deterioration and cross contamination
Solution Approach 1:
The harmful reagent residues and enzyme fragments are extracted and separated from the sample through solid-phase support immobilization. The enzyme is bound to a solid support matrix, allowing the reaction to proceed while enabling easy separation of the enzyme-bound support from the sample solution through filtration or centrifugation, thus removing contamination sources.
Solution Approach 2:
A solid-phase support acts as an intermediary carrier for the enzyme. Instead of using free enzyme solution that mixes with the sample, the enzyme is immobilized on an inert solid support that serves as a mediator, allowing the enzyme to perform its function while remaining physically separable from the sample.
2Object-generated harmful factors
If reusable enzyme-immobilized columns are used, then enzyme residue mixing into sample is reduced, but performance deterioration and cross contamination occur through repeated use
Solution Approach 1:
The invention employs disposable single-use enzyme-immobilized cartridges or columns that are discarded after a single use. This eliminates the performance deterioration and cross-contamination issues associated with reusing columns, as each new cartridge provides a fresh, clean enzyme source without carrying over residues or degraded performance from previous uses.
3Reliability
If immobilized enzyme dispersion is used in disposable system, then cross contamination is suppressed, but reagent residues still mix into the sample
Solution Approach 1:
The harmful reagent residues are extracted and removed from the sample through solid-phase support immobilization. The enzyme is bound to a solid support matrix, allowing the reaction to proceed while enabling easy separation of the enzyme-bound support from the sample solution through filtration or centrifugation, thus removing contamination sources.
4Device complexity
If conventional soluble enzyme method is used, then pretreatment process is simple, but subsequent analysis steps are adversely affected by reagent residues
Solution Approach 1:
A solid-phase support acts as an intermediary carrier for the enzyme. Instead of using free enzyme solution that mixes with the sample, the enzyme is immobilized on an inert solid support that serves as a mediator, allowing the enzyme to perform its function while remaining physically separable from the sample.
Solution Approach 2:
The harmful reagent residues and enzyme fragments are extracted and separated from the sample through solid-phase support immobilization. The enzyme is bound to a solid support matrix, allowing the reaction to proceed while enabling easy separation of the enzyme-bound support from the sample solution through filtration or centrifugation, thus removing contamination sources.
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 the accuracy and efficiency of peptide and protein analysis by minimizing reagent contamination and enabling faster pretreatment processes, while reducing cross-contamination risks through the use of immobilized reagents in a disposable system.
Implementation Method 1
a step A of mixing the sample and a reagent A containing a support and a reducing agent immobilized on the support
Implementation Method 2
a step B of mixing the sample and a reagent B containing a support and an enzyme immobilized on the support
Data Source
AI summary
Objects of the disclosure are to provide a preanalysis treatment method for a sample, which can reduce mixing of a reagent into the sample, and to provide a sample pretreatment system capable of performing the preanalysis treatment method. An aspect of the present embodiment is a preanalysis treatment method for a sample, which includes a step A of mixing the sample and a reagent A containing a support and a reducing agent immobilized on the support, a step B of mixing the sample and a reagent B containing a support and an enzyme immobilized on the support, a step X-A of, after the step A, separating the reagent A and a supernatant from each other, and a step X-B of, after the step B, separating the reagent B and a supernatant from each other.