Chromatography Device with AEX and HIC Layers for Nucleic Acid Purification
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Solution Overview
Problem
Current chromatography devices for RNA and DNA purification require multiple devices and methods, leading to complexity and high buffer consumption, especially when dealing with nucleic acids like lentivirus and AAV, which demand improved efficiency and stability.
Innovation Solution
A chromatography device with a filter housing containing a sequence of anionic exchange (AEX) and hydrophobic interaction (HIC) layers, allowing for a single device to perform multiple purification steps with high flow rates and reduced buffer usage, where nucleic acids are bound and eluted in high and low salt conditions respectively.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple chromatography devices are used for RNA and DNA purification, then purification effectiveness is improved, but device complexity and buffer consumption increase
Solution Approach 1:
The patent combines multiple chromatography functions (anionic exchange and hydrophobic interaction) into a single device with multiple layers. The first porous filter element contains AEX layers for DNA purification while the second porous filter element contains HIC layers for RNA purification, allowing both purification mechanisms to operate simultaneously in one device rather than requiring separate devices.
Solution Approach 2:
The single chromatography device is designed to perform multiple purification functions simultaneously. It can purify both DNA and RNA in the same feed stream through different layers, making the device universal for handling multiple nucleic acid types without requiring separate specialized devices for each purification task.
2Reliability
If multiple chromatography devices are used for purification, then purification effectiveness is improved, but buffer consumption increases
Solution Approach 1:
By merging multiple purification functions into one device, the system eliminates the need for sequential buffer consumption across multiple devices. The high salt buffer used to elute DNA from the AEX layer can be directly reused for the HIC layer purification, significantly reducing overall buffer consumption while maintaining effective purification of both nucleic acid types.
3Ease of operation
If traditional purification methods are used, then process simplicity is maintained, but buffer dilution is required reducing efficiency
Solution Approach 1:
The invention changes the operational parameters by using high salt buffers that enable direct elution without dilution. The high conductivity buffer serves dual purposes: eluting bound DNA from the AEX layer and providing appropriate conditions for HIC layer binding, eliminating the need for buffer dilution steps and improving overall purification efficiency while maintaining operational simplicity.
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 simplifies the purification process, reduces buffer consumption, and enhances the stability of nucleic acids like lentivirus and AAV by using a single device for dual purification actions, achieving high purity without the need for buffer dilution.
Implementation Method 1
the first porous filter element comprises at least one anionic exchange (AEX) layer
Implementation Method 2
binding the nucleic acid to the at least one AEX layer
Implementation Method 3
the second porous filter element comprises at least one hydrophobic interaction (HIC) layer
Implementation Method 4
binding the nucleic acid to the at least one HIC layer
Implementation Method 5
eluting the nucleic acid from the at least one AEX layer in high salt
Implementation Method 6
eluting the nucleic acid in low salt buffer
Data Source
AI summary
A chromatography device is provided comprising a filter housing having an inlet and an outlet and defining a fluid flow path between the inlet and the outlet; a porous filter arranged in the filter housing across the fluid flow path, the filter comprising first porous filter element; and a second porous filter element in contact with the first porous filter element, wherein the first porous filter element comprises at least one anionic exchange (AEX) layer, and the second porous filter element comprises at least one hydrophobic interaction (HIC) layer. A method of purifying nucleic acid using the device is also provided.