Soluble Thrombomodulin Purification via Hydrophobic Interaction Chromatography
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Solution Overview
Problem
Current methods for producing highly-purified soluble thrombomodulin often result in residual host cell proteins, which can cause anaphylactic shock and pose a lethal risk, and existing multi-step column chromatography approaches either fail to further reduce contamination or decrease yield.
Innovation Solution
A method involving filtration through nylon and/or polyethersulfone membranes is employed to produce highly-purified soluble thrombomodulin with reduced host cell protein contamination, achieving a concentration of less than 10 ng per 10,000 U, thereby enhancing safety and efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If multi-step column chromatography is used to purify soluble thrombomodulin, then manufacturing precision is improved, but productivity deteriorates due to decreased yield
Solution Approach 1:
The patent changes the purification method from multi-step column chromatography to a single-step hydrophobic interaction chromatography method by altering the purification parameters and conditions, achieving both high purification level and maintained yield
Solution Approach 2:
The patent extracts and eliminates the need for multiple chromatography steps by using a single hydrophobic interaction chromatography step that simultaneously achieves both purification and maintains yield, removing the trade-off between precision and productivity
2Ease of manufacture
If existing purification methods are used, then manufacturing process is simple, but object-affected harmful factors increase due to residual host cell proteins causing anaphylactic shock
Solution Approach 1:
The patent changes the chromatography method to hydrophobic interaction chromatography with specific buffer conditions (pH 7.0-9.0, 0.1-1.0 M salt concentration) that simultaneously maintain process simplicity and achieve extremely low host cell protein contamination levels
Solution Approach 2:
The patent uses a single-use purification approach with hydrophobic interaction chromatography that replaces complex multi-step procedures, maintaining ease of manufacture while achieving superior purification that eliminates harmful host cell protein residues
3Device complexity
If existing purification methods are used, then device complexity is low, but manufacturing precision deteriorates due to inability to further reduce contamination
Solution Approach 1:
The patent changes the chromatography parameters to hydrophobic interaction conditions that enable extremely high purification levels with a single step, achieving manufacturing precision of less than 10 ng HCP per 10,000 U without increasing device complexity
Solution Approach 2:
The hydrophobic interaction chromatography method serves multiple functions simultaneously: it purifies soluble thrombomodulin, removes host cell proteins, and maintains yield, achieving high manufacturing precision without requiring complex multi-step systems
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 method effectively reduces host cell protein contamination to a safe level, ensuring the purified thrombomodulin has a purity of 99% or higher and maintains its biological activities, such as thrombin binding, Protein C activation, and anti-inflammatory effects.
Implementation Method 1
passing a solution containing soluble thrombomodulin through a nylon and/or polyethersulfone filtration membrane
Data Source
AI summary
Highly-purified soluble thrombomodulin which has a content of host cell-originated proteins being in a ratio of less than 10 ng of the proteins per 10,000 U of the soluble thrombomodulin, wherein the soluble thrombomodulin is produced by a transformant cell obtained by transfecting a host cell with a DNA containing a nucleotide sequence encoding the soluble thrombomodulin.