Size Exclusion Matrix for Protein Separation
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Solution Overview
Problem
Current sample preparation techniques struggle to effectively separate larger proteins from smaller proteins and recover low concentration and/or low abundance proteins, especially in small and large column formats, leading to inefficient downstream processing.
Innovation Solution
A matrix comprising a porous size exclusion support with a molecular weight cut-off (MWCO) greater than 30,000 Da, made from a reaction product of 2-hydroxyethyl cellulose and epichlorohydrin, and optionally modified with an amine-containing moiety, is used for separating small molecules from large molecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional sample preparation techniques are used, then separation of small molecules from large molecules can be achieved, but the separation efficiency is insufficient and low concentration proteins cannot be effectively recovered
Solution Approach 1:
The patent employs a porous size exclusion support with specifically engineered pore sizes that enable differential penetration of molecules based on their size. The porous structure allows small molecules to enter and be retained while excluding larger proteins, achieving both high separation efficiency and effective recovery of low concentration proteins through the size-based filtration mechanism
Solution Approach 2:
The patent modifies the physical and chemical parameters of the support material by incorporating specific functional groups and adjusting pore size distribution. These parameter changes optimize the interaction between the matrix and target molecules, enhancing both separation precision and the ability to recover low concentration proteins that would otherwise be lost in conventional methods
2Productivity
If conventional matrices are used for separation, then basic filtration can be performed, but superior separation and reduced processing time are not achieved
Solution Approach 1:
The patent utilizes a composite matrix structure combining size exclusion support with functional groups that provide both rapid filtration capabilities and high-selectivity separation. This composite material architecture enables simultaneous achievement of fast processing speeds through efficient flow characteristics and superior separation quality through multifunctional interaction mechanisms
Solution Approach 2:
The designed matrix exhibits multi-functionality by simultaneously providing size-based exclusion, adsorption capabilities through functional groups, and flow optimization. This universal design allows a single matrix to perform multiple separation functions, achieving both rapid processing and high-quality separation without requiring multiple sequential steps
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 described matrix enables superior separation of small molecules from large molecules in any column size, reducing time and expense, and facilitates the recovery of low concentration proteins, thereby enhancing downstream processing efficiency.
Implementation Method 1
a porous size exclusion support having a molecular weight cut-off (MWCO) greater than 30,000 Da (30 kDa)
Data Source
AI summary
Disclosed herein are aspects of a matrix, typically comprising a rigid resin, for purifying samples, such as by separating one or more small molecules from one or more large molecules in a sample. Also disclosed is a method of making the matrix, a method of using the matrix, systems comprising the matrix, and kit aspects. Disclosed aspects comprising a porous size exclusion support and an amine-containing moiety can be used in small and large spin column formats and can be equilibrated with a wide range of buffers.