Antibody Flow-Through Chromatography Using Tris Conductivity Adjustment
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Solution Overview
Problem
Existing antibody purification methods using multimodal ion exchange chromatography in flow-through mode suffer from low yield and inefficient removal of aggregates and impurities, particularly when multiple chromatography steps are involved, leading to prolonged processing times and potential protein instability.
Innovation Solution
The method involves pre-conditioning the antibody sample with Tris to adjust conductivity prior to loading the chromatography column, eliminating the use of NaCl, thereby enhancing antibody yield and reducing aggregates and impurities during multimodal anion exchange chromatography.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If multiple chromatography steps are used to remove aggregates and impurities, then purity is improved, but processing time increases and protein stability deteriorates
Solution Approach 1:
The patent combines multiple chromatography functions into a single multimodal chromatography step that simultaneously performs capture, removal of aggregates, and removal of impurities. This integration reduces the number of sequential steps from multiple separate chromatography operations to one consolidated process, thereby reducing processing time while maintaining purity requirements.
Solution Approach 2:
The multimodal chromatography medium is designed to perform multiple purification functions simultaneously - capturing antibodies, removing aggregates, and eliminating various impurities (HCPs, nucleic acids, viruses, medium components). This multi-functional approach replaces what would traditionally require multiple specialized chromatography steps, improving efficiency and reducing processing time.
2Ease of manufacture
If conductivity is adjusted with NaCl prior to IEX step, then standard purification protocol is followed, but antibody yield decreases
Solution Approach 1:
The patent changes the chemical parameter used for conductivity adjustment from NaCl to Tris buffer. This substitution modifies the ionic composition of the sample before IEX chromatography, creating favorable conditions for antibody flow-through while maintaining appropriate conductivity levels. The Tris buffer provides conductivity adjustment without the adverse effects of NaCl on antibody binding and yield.
Solution Approach 2:
Tris buffer serves as an intermediary substance that performs the conductivity adjustment function without directly interfering with antibody-chromatography medium interactions. Unlike NaCl which can compete for binding sites, Tris acts as a neutral mediator that establishes proper ionic strength and pH conditions, enabling high antibody yield while maintaining protocol structure.
3Productivity
If Tris is added to adjust conductivity, then antibody yield increases, but monomer content decreases slightly
Solution Approach 1:
The patent extracts or removes aggregates and impurities through the multimodal chromatography step, allowing the antibody monomers to pass through in the flow-through fraction. The slight decrease in monomer content is acceptable because the overall process achieves high purity by removing contaminants, and the yield improvement indicates successful separation of monomers from aggregates and impurities.
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 results in a 5% higher antibody yield and slightly lower monomer content while maintaining purity, demonstrating improved efficiency and effectiveness in antibody purification.
Implementation Method 1
pre-conditioning the antibody sample with Tris to adjust conductivity prior to loading the chromatography column
Implementation Method 2
multimodal anion exchange chromatography in flow-through mode
Implementation Method 3
The mode of operation also plays an important part for successful multimodal chromatography. In flow-through (FT) mode, for example, the pH of the sample and buffer is selected to modify the charge of the antibody or the chromatography medium such that the antibody will not bind but will rather flow through the column
Data Source
AI summary
The present invention relates to a method for increasing antibody yield during antibody purification from a sample by ion exchange chromatography in flow through mode by pre-conditioning the sample with Tris without the use of NaCl to adjust the conductivity.