Multidimensional Liquid Chromatography System for Protein Separation

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current multidimensional liquid chromatography (mD-LC) systems for protein separation face limitations such as low sample injection capacity, compatibility issues between mobile phases, and inability to achieve high-purity protein separation on an industrial scale, particularly due to limitations in chromatographic column design and operational complexity.

Innovation Solution

A multidimensional liquid chromatography separation system that includes a mobile phase tank, independent infusion devices, sample injection systems, a chromatographic column switching unit, and a collect-reserve device, allowing for on-line two-dimensional or multi-dimensional separation with adjustable flow rates and eluent concentrations to manage protein retention and separation, enabling higher sample throughput and purity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional chromatographic columns are used for protein separation, then the separation can be performed with simple equipment, but the separation purity and resolution are insufficient for complex protein mixtures

Engineering Contradiction:
Improveseparation purityVSAvoidchromatography system complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention divides the chromatography process into multiple dimensions (e.g., first dimension and second dimension separations) using different separation mechanisms. Each dimension uses a dedicated chromatographic column with specific stationary phase properties, allowing complex protein mixtures to be separated step-by-step with high resolution and purity.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention transitions from single-dimensional chromatography to multi-dimensional chromatography by adding temporal and spatial dimensions. Fractions from the first dimension are collected and re-injected into the second dimension column, creating a two-dimensional separation space that dramatically improves resolution and purity for complex samples.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

2Manufacturing precision

If multiple chromatographic columns are connected in parallel for multidimensional separation, then the separation capability is improved, but the system complexity and operational difficulty increase significantly

Engineering Contradiction:
Improveseparation resolutionVSAvoidoperational simplicity
Core Design Contradiction:
Manufacturing precisionVSEase of operation

Solution Approach 1:

The invention merges multiple chromatographic columns and separation dimensions into an integrated system with unified control. The first and second infusion devices, injector devices, and collect-reserve devices work together as a coordinated whole, automatically managing the complex multi-dimensional separation process and simplifying operation despite the advanced separation capability.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The invention introduces a collect-reserve device as an intermediary between the first and second dimension separations. This device collects fractions from the first dimension, stores them, and re-injects them into the second dimension column, acting as a mediator that enables the transition between dimensions while managing system complexity.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Ease of operation

If only a small fraction of collected samples is injected into the second column, then the system operation is simplified, but the sample recovery rate decreases

Engineering Contradiction:
Improvesystem operabilityVSAvoidsample recovery rate
Core Design Contradiction:
Ease of operationVSQuantity of substance

Solution Approach 1:

The invention performs preliminary fraction collection and concentration in the collect-reserve device before second dimension injection. This preliminary action allows the system to optimize injection volumes for each fraction, maximizing sample recovery while maintaining operational simplicity and separation quality.

Inventive Principle:
Principle #10Preliminary action

4Manufacturing precision

If different mobile phases are used for different chromatography patterns, then the separation selectivity is improved, but the compatibility problems between mobile phases arise

Engineering Contradiction:
Improveseparation selectivityVSAvoidmobile phase compatibility
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The invention segments the mobile phase system into dimension-specific phases, with the first infusion device providing mobile phase for the first dimension and the second infusion device providing mobile phase for the second dimension. Each mobile phase is optimized for its specific separation mechanism, maintaining high selectivity while avoiding compatibility issues through separate delivery systems.

Inventive Principle:
Principle #1Segmentation

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 system enhances protein separation efficiency, increases sample recovery, and reduces contamination and operational time, enabling rapid analysis and large-scale preparation of proteins with improved detection sensitivity and cost-effectiveness.

Implementation Method 1

chromatographic column (7) or chromatographic cakes (14)... separation device, which includes chromatographic column switching unit (6-1, 6-2) and n (n≥1) chromatographic columns (7) or chromatographic cakes (14)

Methodology Applied
Scientific EffectAdsorption: Adsorption

Implementation Method 2

the first and second infusion device (3, 9), which works independently. They take mobile phase for liquid chromatography separation from the mobile phase tank (2-1 to 2-4) and then, the mobile phase transfers to mD-LC separation system flow path

Methodology Applied
Scientific EffectPressure-driven flow: Pressure Gradient

Implementation Method 3

detection device (13)

Methodology Applied
Scientific EffectUV absorption: Absorption (EM radiation)

Data Source

PatentEP2891881B1Multidimensional liquid chromatography separation system and separation method for protein separation
Publication Date: 2023.07.26 XIAN AOLAN SCI & TECH
  • EP2891881B1 patent drawingFigure 1A~1B
  • EP2891881B1 patent drawingFigure 1C~1D
  • EP2891881B1 patent drawingFigure 1E~1F

AI summary

A multidimensional liquid chromatography separation system and separation method for protein separation. The multidimensional liquid chromatography separation system for protein separation comprises a mobile-phase storage tank (2), a first liquid transfer device (3, 1-2), a second liquid transfer device (9, 1-1), a first sample introduction device (4-1, 4-2), a second sample introduction device (12), a separation device (7), a collection and storage device (10), at least two drainage devices (7-1, 7-2, 7-3) and a flow path switching device (11). When the liquid chromatography separation of each dimension is conducted, the collection and storage device collects target middle distillates needing to conduct the next step of liquid chromatography separation and stores same in a collection and storage device. During the liquid chromatography separation of the second dimension or more dimensions, all the target middle distillates from the collection and storage device are mixed with mobile phases through a sample introduction mixer of the second sample introduction device, and the amount of the mobile phases conveyed by same is adjusted and measured using the first liquid transfer device and the second liquid transfer device, so as to enable the eluent concentration in the sample introduction mixture to be lower than the critical migration eluent concentration of all the target proteins in the target middle distillates which need to be preserved in the separation of this dimension.