Factor X Purification via Immobilized Metal Ion Affinity Chromatography
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Solution Overview
Problem
Current treatments for factor X deficiency, such as plasma and prothrombin complex concentrates, are inefficient, require large volumes, pose risks of volume overload, thrombotic side effects, and have variable dosing due to high prothrombin contamination, necessitating a more effective method for separating factor X from prothrombin.
Innovation Solution
The method employs immobilized metal ion affinity chromatography (IMAC) followed by anion exchange chromatography to selectively separate and purify factor X, allowing for the collection of factor X-enriched fractions while discarding prothrombin-rich fractions, thereby reducing thrombotic risks and achieving higher purity and concentration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If prothrombin complex concentrates are used to treat factor X deficiency, then factor X delivery is improved, but prothrombin contamination increases leading to thrombotic risks
Solution Approach 1:
The patent applies extraction by removing prothrombin from the prothrombin complex concentrate through chromatography techniques. The process selectively extracts prothrombin while retaining factor X, thereby delivering factor X therapy while eliminating the harmful prothrombin component that causes thrombotic risks.
Solution Approach 2:
The patent utilizes parameter changes in chromatography conditions (pH, ionic strength, temperature) to alter the binding affinity of prothrombin and factor X to the chromatography medium. By adjusting these parameters, prothrombin is selectively bound and removed while factor X remains in solution, resolving the contradiction between factor X delivery and thrombotic risk reduction.
2Quantity of substance
If plasma is used for factor X replacement, then factor X is delivered to patients, but large infusion volumes are required causing volume overload
Solution Approach 1:
The patent extracts factor X from plasma through chromatography, concentrating it into a small-volume pharmaceutical composition. This extraction process removes factor X from the large-volume plasma matrix, enabling delivery of therapeutic factor X doses in minimal infusion volumes that avoid volume overload.
Solution Approach 2:
The patent creates a composite pharmaceutical composition containing factor X combined with stabilizing excipients and buffering agents. This composite formulation allows factor X to be delivered in concentrated, stable, small-volume form rather than requiring large volumes of native plasma.
3Manufacturing precision
If multiple chromatography steps are used to purify factor X, then prothrombin removal is improved, but process complexity increases
Solution Approach 1:
The patent segments the purification process into distinct chromatography steps, each targeting specific impurities. The first chromatography step removes prothrombin, while subsequent steps remove other contaminants. This segmentation allows systematic removal of prothrombin with controlled complexity at each stage rather than attempting single-step removal.
Solution Approach 2:
The patent uses chromatography media as intermediary substances that selectively bind prothrombin while allowing factor X to pass through. These intermediary chromatography resins mediate the separation process, enabling high-purity prothrombin removal without requiring complex mechanical or chemical processing equipment.
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 process enables the production of highly purified factor X with reduced prothrombin contamination, allowing for a more stable and effective pharmaceutical formulation that can be stored at higher temperatures, incorporating multiple virus reduction steps for safety, and minimizing thrombotic side effects during clinical use.
Implementation Method 1
a) adsorbing the starting material onto an immobilised metal ion affinity chromatography substrate
Implementation Method 2
The second portion of the eluate may then be subjected to a further purification step using anion exchange chromatography
Data Source
AI summary
Methods for preparing Factor X, activated Factor X, inactivated factor X and inactivated factor Xa, compositions comprising Factor X and Factor Xa, inactivated Factor X and inactivated Factor Xa and methods of medical treatment using Factor X, Factor Xa, activated Factor X and inactivated Factor Xa are disclosed. The preparation methods comprise a chromatography step using an immobilised metal ion affinity chromatography substrate.