Protein C S Concentrate Plasma Purification
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Solution Overview
Problem
Current methods for treating sepsis with Protein C concentrates lack safety due to high hemorrhagic risk, especially in pediatric patients, and do not effectively utilize Protein S, which could provide additional anticoagulant and anti-inflammatory benefits, while existing Protein C concentrates lack Protein S and are costly.
Innovation Solution
A process is developed to purify a concentrate of Protein C containing Protein S from human plasma, involving multiple chromatographic steps using DEAE-Sepharose, weak anion-exchange resin, and hydroxyapatite, followed by viral inactivation and nanofiltration, to produce a safe and cost-effective product for parenteral administration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If activated recombinant Protein C is used to treat sepsis, then anticoagulant effect is achieved, but hemorrhagic risk increases significantly
Solution Approach 1:
The patent combines Protein C and Protein S into a single concentrate product, utilizing their synergistic anticoagulant effects. Protein S enhances the anticoagulant activity of Protein C while reducing the required dose, thereby achieving therapeutic effect with lower exposure and reduced hemorrhagic risk.
Solution Approach 2:
The patent uses multiple chromatographic steps (DEAE-Sepharose, weak anion-exchange, hydroxyapatite) to precisely control the composition, concentration, and purity of Protein C and Protein S in the concentrate, optimizing the therapeutic index by maintaining appropriate protein ratios and eliminating contaminants.
2Manufacturing precision
If Protein C concentrate is purified using existing methods, then Protein C is obtained, but Protein S is lost or not utilized
Solution Approach 1:
The chromatographic process is designed to simultaneously purify and concentrate both Protein C and Protein S from plasma. The multi-step chromatography system serves multiple functions: removing contaminants, separating proteins by specific binding properties, and concentrating both target proteins in the final eluate.
Solution Approach 2:
The patent uses specific chromatographic resins (DEAE-Sepharose, weak anion-exchange resin, hydroxyapatite) as intermediary substances that selectively bind and release Protein C and Protein S based on their biochemical properties, enabling their co-purification while excluding other plasma proteins.
3Reliability
If activated recombinant Protein C is produced, then therapeutic effect is achieved, but production cost increases
Solution Approach 1:
The patent uses plasma-derived proteins instead of expensive recombinant production. The starting material (human plasma or plasma-derived fractions) is more cost-effective than establishing and maintaining complex recombinant protein production systems, while still providing the necessary therapeutic effect.
Solution Approach 2:
The invention recovers valuable Protein C and Protein S from plasma processing waste streams or underutilized fractions, transforming low-value byproducts into high-value therapeutic products and reducing overall production costs.
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 process results in a safe and effective Protein C and Protein S concentrate that reduces hemorrhagic risks, provides synergistic anticoagulant and anti-inflammatory effects, and is more cost-effective than activated recombinant Protein C, suitable for patients where traditional treatments are contraindicated.
Implementation Method 1
the fraction collected after the second wash of DEAE-Sepharose resin used for obtaining the purified Prothrombin Complex (PTC) starting from frozen human plasma
Implementation Method 2
the product derived from the previous step is subjected to chromatography on weak anion-exchange resin and the pH of the eluate containing the proteins of interest is adjusted
Implementation Method 3
a second chromatographic step is carried out on a resin which selectively binds Protein C and Protein S, wherein said resin is hydroxyapatite
Implementation Method 4
the purified protein mixture obtained in this way is dialyzed with a suitable formulation buffer, then prefiltered, nanofiltered and lyophilised
Data Source
Figure 1
AI summary
Described is a concentrate of Protein C also comprising Protein S, a method for its extraction from human plasma as well as its use in the treatment of congenital and acquired deficiencies of Protein C and/or Protein S and in the treatment of sepsis.