Bacteriophage Purification Using Selective Permeable Membranes
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Solution Overview
Problem
Current bacteriophage purification methods are inefficient, leading to variable results in antibacterial activity and high contamination levels, particularly with bacterial endotoxins, which limits their effectiveness and scalability for phage therapy and pharmaceutical applications.
Innovation Solution
A continuous purification method using selective permeable membranes with specific cut-offs, combined with enzymatic treatments and gentle washing buffers, to retain bacteriophages while removing low molecular weight molecules and contaminants, resulting in high-titer, low-endotoxin preparations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If traditional purification methods (ultracentrifugation, dialysis, gel filtration, ultrafiltration) are used to remove endotoxins and contaminants, then bacteriophage purification is achieved, but the process becomes complex and time-consuming with variable results
Solution Approach 1:
The patent extracts and removes harmful contaminants (endotoxins, bacterial proteins, lipids) from the bacteriophage preparation using selective permeable membranes with specific cut-off sizes. The method systematically separates contaminants based on their molecular weight, allowing retention of bacteriophages while eliminating low molecular weight impurities through controlled filtration processes.
Solution Approach 2:
The patent introduces selective permeable membranes as intermediary elements between the bacteriophage suspension and the purification system. These membranes act as mediators that selectively allow certain molecules to pass while blocking others, enabling controlled purification without requiring complex multi-step procedures.
2Reliability
If multiple purification steps (centrifugation, filtration, molecular filtration) are performed to reduce endotoxin levels, then bacteriophage activity is maintained, but the process time and resource consumption increase
Solution Approach 1:
The patent implements continuous purification processes where bacteriophage suspension flows continuously through selective permeable membranes. The system maintains continuous separation and concentration without interruption, eliminating the need for repeated batch processing steps and significantly reducing total process time while maintaining high bacteriophage activity.
Solution Approach 2:
The selective permeable membranes perform multiple purification functions simultaneously - separating contaminants, concentrating bacteriophages, and reducing endotoxin levels - without requiring external intervention or multiple specialized steps. The system serves multiple purposes through a single integrated mechanism.
3Quantity of substance
If conventional purification methods are used, then some bacteriophage preparation is obtained, but contamination with bacterial endotoxins and toxins remains high
Solution Approach 1:
The patent applies selective permeable membranes with specific local properties (defined cut-off sizes) to target specific contaminants. Different membrane sections or stages are optimized for removing specific types of contaminants based on their molecular size, enabling precise removal of endotoxins and low molecular weight toxins while preserving bacteriophages.
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 method achieves consistent bacteriophage purification with high antibacterial activity, significantly reducing bacterial contaminants and endotoxin levels, enabling effective phage therapy and pharmaceutical applications on an industrial scale.
Implementation Method 1
The purification and concentration is made in a continuous manner using selective permeable membrane which retains bacteriophages but not low molecular mass molecules
Implementation Method 2
The bacteriophage particles can be then concentrated using ultrafiltration on a membrane with cut off from 1000 to 100000 Da, advantageously from 1000 to 10000 Da
Data Source
AI summary
A method of preparation of purified bacteriophage with increased antibacterial activity, in which from bacterial lysate of phages is obtained, advantageously in the presence of lysozyme, chelating factor and detergent, in a continuous manner with ultrafiltration on membranes, the phage containing high molecular mass preparation, devoid of bacterial cell wall and other contaminants, free of toxins and endotoxins, active in tests of bacterial lysis, which is characterized by chromatography HPLC, In SDS-PAGE electrophoresis, immunoblotting, biological tests of bacterial lysis, and is dedicated for phage therapy of bacterial infections and tumors and for production of phage deriving pharmaceutical preparations.