Extraction Column Low Pore Volume Frits for Biomolecule Purification
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Solution Overview
Problem
Current methods for purifying and concentrating biomolecules from small volume samples often result in sample loss and inefficiency due to the need for removing interfering contaminants, particularly in proteomics and other analytical techniques.
Innovation Solution
The development of extraction columns with a packed bed of gel-type resin beads, utilizing a bed of extraction media with a low volume and low pore volume frits, which allows for efficient adsorption, desorption, and elution of analytes using a small volume of desorption solvent, minimizing sample loss and maximizing enrichment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional solid phase extraction methods are used for purifying and concentrating biomolecules from small volume samples, then purification and concentration can be achieved, but sample loss occurs and interfering contaminants are not effectively removed
Solution Approach 1:
The patent employs porous frits with specific pore volumes (top frit pore volume ≤ 10% of interstitial volume, bottom frit pore volume ≤ 20% of interstitial volume) to enable effective purification while minimizing sample loss. The porous structure allows selective retention of analytes while permitting flow of solvents and removal of contaminants, resolving the contradiction between purification effectiveness and sample loss.
2Quantity of substance
If larger beds of extraction media are used to improve purification capacity, then more analyte can be processed, but sample loss increases and enrichment efficiency decreases
Solution Approach 1:
The patent optimizes the pore volume parameters of the frits (top frit pore volume ≤ 10% of interstitial volume, bottom frit pore volume ≤ 20% of interstitial volume) to achieve effective purification with minimal sample loss. By carefully controlling these physical parameters, the system maintains high enrichment factors even when processing larger quantities of analyte, resolving the contradiction between processing capacity and sample loss.
3Productivity
If conventional frits with high pore volume are used, then solvent flow is facilitated, but analyte retention is reduced and enrichment efficiency decreases
Solution Approach 1:
The patent specifies precise pore volume constraints for the frits (top frit pore volume ≤ 10% of interstitial volume, bottom frit pore volume ≤ 20% of interstitial volume) to balance solvent flow and analyte retention. This controlled porosity allows adequate solvent flow rate while maintaining high enrichment factors, resolving the contradiction between productivity and manufacturing 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
This approach enables effective purification and concentration of biomolecules with high enrichment factors, reducing sample loss and maintaining the integrity of the analytes, suitable for proteomics and other analytical applications.
Implementation Method 1
a packed bed of extraction media, e.g., a bed of gel-type beads derivatized with a group having an affinity for an analyte of interest
Implementation Method 2
the top frit having a low pore volume, wherein the top frit, bottom frit, and column body define an extraction media chamber
Data Source
AI summary
The invention provides extraction columns for the purification of an analyte (e.g., a biological macromolecule, such as a peptide, protein or nucleic acid) from a sample solution, as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the column between two frits. In some embodiments, the extraction columns employ modified pipette tips as column bodies. In some embodiments, the invention provides methods of storing columns in a wet state.