Affinity Chromatography Column for Weak Separation of Biological Samples
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Solution Overview
Problem
Current methods for detecting target compounds in biological samples, such as blood serum, face challenges due to low concentrations and interference from proteins, requiring multiple separation steps and dilution, which complicates the analysis process.
Innovation Solution
A method utilizing an affinity column with a high density of preselected binding ligands for weak affinity separation, allowing direct analysis of target compounds from whole biological samples via mass spectrometry, enabling efficient separation and detection without prior protein removal or dilution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If current separation methods are used to remove proteins from serum, then protein interference is reduced, but the analysis process becomes more complex and requires multiple separation steps
Solution Approach 1:
The invention combines protein removal and target compound separation into a single affinity chromatography step. The affinity column is designed with specific ligands that simultaneously bind and remove proteins while allowing target compounds to pass through, eliminating the need for multiple separate separation steps and reducing overall process complexity
Solution Approach 2:
The affinity column serves multiple functions simultaneously: it acts as both a protein removal medium and a separation medium for target compounds. This multi-functional approach allows a single step to achieve what previously required multiple sequential operations, simplifying the analytical workflow
2Productivity
If target compounds are detected in undiluted biological samples, then analysis efficiency is improved, but protein interference and matrix effects increase
Solution Approach 1:
The invention extracts and removes proteins and interfering matrix components from undiluted biological samples using affinity chromatography before detection. The affinity column selectively binds proteins while allowing target compounds to elute in a cleaned-up form, enabling direct analysis of undiluted samples without interference
Solution Approach 2:
The affinity column acts as an intermediary between the complex biological sample and the detection system. It mediates the interaction by selectively removing proteins and interfering substances, allowing the detector to analyze target compounds in undiluted samples without being affected by matrix components
3Manufacturing precision
If affinity column has high density of binding ligands, then separation capability is improved, but column capacity for processing large sample volumes is reduced
Solution Approach 1:
The invention applies local quality by creating zones of different ligand densities within the affinity column. High ligand density regions provide excellent separation resolution for target compounds, while lower density regions maintain higher binding capacity for processing large sample volumes. This spatial variation in ligand distribution optimizes both separation performance and processing capacity
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 allows for the effective separation and detection of target compounds, such as steroids, in undiluted biological samples, improving analysis efficiency and reducing the need for multiple separation steps, while maintaining column throughput.
Implementation Method 1
delivering a biological sample through an affinity column, the affinity column having a binding ligand coupled to a stationary structural support, wherein the affinity column has a high density of the binding ligand per the stationary structural support and wherein the binding ligand has been preselected to cause weak affinity separation zonal retardation of the target compound from the biological sample
Data Source
AI summary
The present invention provides methods for analyzing a target compound from a biological sample. In one aspect, a method for analyzing a target compound in a biological sample can comprise delivering a biological sample through an affinity column, the affinity column having a binding ligand coupled to a stationary structural support, wherein the affinity column has a high density of the binding ligand per the stationary structural support and wherein the binding ligand has been preselected to cause weak affinity separation zonal retardation of the target compound from the biological sample forming a target compound fraction and a biological sample fraction and detecting the target compound by mass spectrometry.