Virus-Like Particle Purification via Ultrafiltration Membrane

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Solution Overview

Problem

Current methods for purifying virus-like particles (VLPs) are inadequate for large-scale production, particularly in meeting GMP compliance and are complex, leading to impurities and inefficiencies.

Innovation Solution

A method involving filtration through a membrane with an exclusion limit of 30 kDa to 1500 kDa, allowing efficient separation and concentration of VLPs, maintaining their native form, and enabling buffer exchange, thus improving purity and scalability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If traditional purification methods (density centrifugation, chromatography) are used, then VLP purity is improved, but process complexity and manufacturing cost increase significantly

Engineering Contradiction:
ImproveVLP purityVSAvoidprocess complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention extracts and utilizes the size difference between VLPs and impurities by applying ultrafiltration membrane technology. The membrane selectively retains VLPs while allowing smaller impurities to pass through, achieving purification without complex chromatography or density gradient centrifugation steps.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention replaces complex mechanical purification systems (centrifugation equipment, chromatography columns) with a simpler ultrafiltration membrane system. This substitution maintains purification effectiveness while dramatically reducing process complexity and equipment requirements.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Manufacturing precision

If multiple purification steps are used, then VLP purity is improved, but production time and energy consumption increase

Engineering Contradiction:
ImproveVLP purityVSAvoidproduction time
Core Design Contradiction:
Manufacturing precisionVSLoss of time

Solution Approach 1:

The invention merges multiple purification functions (filtration, concentration, buffer exchange) into a single ultrafiltration step. This consolidation achieves the same purification效果 as multiple sequential steps while significantly reducing production time and energy consumption.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The ultrafiltration membrane system performs multiple functions simultaneously: it filters impurities, concentrates VLPs, and enables buffer exchange. This multi-functionality eliminates the need for separate operation steps, reducing overall production time.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Ease of manufacture

If conventional filtration is used, then process simplicity is improved, but VLP recovery efficiency decreases due to loss through the filter

Engineering Contradiction:
Improveprocess simplicityVSAvoidVLP recovery efficiency
Core Design Contradiction:
Ease of manufactureVSLoss of substance

Solution Approach 1:

The invention changes the key parameter of filter pore size to be smaller than VLPs but larger than impurities. This parameter optimization ensures that VLPs are retained on the membrane while impurities pass through, achieving both process simplicity and high VLP recovery efficiency.

Inventive Principle:
Principle #35Parameter changes

4Manufacturing precision

If complex chromatography steps are used, then VLP purity is improved, but scalability to industrial production is limited

Engineering Contradiction:
ImproveVLP purityVSAvoidscalability
Core Design Contradiction:
Manufacturing precisionVSProductivity

Solution Approach 1:

The invention extracts the essential purification function from complex chromatography systems and implements it through ultrafiltration membrane technology. This extraction enables the purification process to be scaled to industrial production levels while maintaining high VLP purity.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention replaces complex chromatography mechanical systems with a scalable ultrafiltration membrane system. This substitution maintains purification effectiveness while enabling easy scaling from laboratory to industrial production.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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 high-purity VLPs suitable for GMP-compliant large-scale production by efficiently separating VLPs from impurities, concentrating the VLPs, and maintaining their native form, enhancing subsequent processing efficiency.

Implementation Method 1

a VLP-containing composition is filtered through a filter medium, in particular a membrane, the filter medium having an exclusion limit (Molecular Weight Cut off (MWCO) of more than 30 kDa to 1500 kDa

Methodology Applied
Scientific EffectUltrafiltration: Semipermeable Membrane

Data Source

PatentEP2739725B1Method for purifying virus-like particles
Publication Date: 2019.11.06 LIFE SCI INKUBATOR BETRIEBS
  • EP2739725B1 patent drawingFigure 1
  • EP2739725B1 patent drawingFigure 2
  • EP2739725B1 patent drawingFigure 3

AI summary

The invention relates to a method for purifying compositions containing virus-like particles (VLP), wherein a VLP-containing composition is filtered through a filter medium, in particular through a membrane, having a molecular weight cut off (MWCO) of more than 30 kDa, and the cell culture supernatant of VLP-expressing cells is used as a VLP-containing composition. The invention further relates to a VLP-containing composition which can be produced by this method.