Virus Stabilization via PVP and Amino Acids in Freeze-Drying

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Solution Overview

Problem

Biologically active materials, such as virus-based vaccines, face challenges in maintaining structural integrity during storage and shipping due to thermal degradation, especially in developing nations with limited infrastructure, and existing stabilization methods are costly, inconvenient, or pose safety risks.

Innovation Solution

A freeze-dried formulation comprising a poxvirus-based material with polyvinylpyrrolidone, sucrose, arginine, glutamate, and phosphate salts, which maintains stability at refrigerated temperatures, reducing virus titer loss and preserving biological activity during storage and shipping.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Stability of the object's composition

If ultra-low temperature storage (below 0°C) is used to maintain viral structural integrity, then virus stability is improved, but storage cost and operational complexity increase significantly

Engineering Contradiction:
Improveviral structural integrityVSAvoidstorage infrastructure
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The invention changes the storage temperature parameter from ultra-low (below 0°C) to refrigerated (4°C to 8°C) conditions. This parameter change is enabled by the formulation containing specific stabilizing agents (polymer, disaccharide, amino acids, and salts) that protect the virus at higher temperatures, thus resolving the contradiction between maintaining viral integrity and reducing storage infrastructure requirements

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The formulation acts as an intermediary protective medium between the virus and harsh storage conditions. The stabilizing agents (polymer, disaccharide, amino acids, and salts) serve as intermediaries that absorb thermal stress and prevent direct thermal degradation of the viral structure, allowing storage at refrigerated temperatures instead of ultra-low temperatures

Inventive Principle:
Principle #24Intermediary (Mediator)

2Stability of the object's composition

If animal or human origin additives (albumin, peptone, gelatine, haemaccel) are used to stabilize virus, then virus stability is improved, but safety risks and cost increase

Engineering Contradiction:
Improvevirus stabilityVSAvoidallergic reactions and contamination risks
Core Design Contradiction:
Stability of the object's compositionVSObject-affected harmful factors

Solution Approach 1:

The invention replaces expensive and risky animal/human origin additives with synthetic or plant-based alternatives (polymer, disaccharide, amino acids, and salts). These alternative stabilizing agents are safer, cheaper, and do not carry the risks of allergic reactions or contamination associated with animal and human products, thus resolving the contradiction between virus stability and safety

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The invention creates a simplified, safer model of the stabilization function. Instead of using complex biological additives (albumin, peptone, gelatine, haemaccel), the formulation uses a streamlined combination of polymer, disaccharide, amino acids, and salts that achieves the same stabilizing effect without the harmful associations, effectively copying the protective function with safer components

Inventive Principle:
Principle #26Copying

3Stability of the object's composition

If freeze-drying process is applied to dry virus-based material, then storage stability at refrigerated conditions is improved, but virus titer loss occurs during the drying process

Engineering Contradiction:
Improvestorage stability at 4°C to 8°CVSAvoidviral titer
Core Design Contradiction:
Stability of the object's compositionVSLoss of substance

Solution Approach 1:

The formulation provides beforehand cushioning by incorporating stabilizing agents (polymer, disaccharide, amino acids, and salts) that protect the virus during the freeze-drying process. These agents act as protective cushions that absorb the stresses of concentration, precipitation, and pH extremes encountered during drying, thereby reducing viral titer loss while maintaining storage stability

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

Solution Approach 2:

The stabilizing agents serve as intermediaries during the freeze-drying process, mediating between the virus and the harsh drying conditions. The polymer, disaccharide, amino acids, and salts form a protective matrix that facilitates water removal while protecting viral particles from direct exposure to concentrated salts, pH extremes, and mechanical stress, thus reducing titer loss

Inventive Principle:
Principle #24Intermediary (Mediator)

4Quantity of substance

If freeze-drying process is applied to concentrate viral material, then storage concentration is improved, but manufacturing time and process complexity increase

Engineering Contradiction:
Improvevirus concentrationVSAvoiddrying process time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

The invention changes the formulation parameters by incorporating specific stabilizing agents (polymer, disaccharide, amino acids, and salts) that enable effective concentration and drying at lower temperatures and shorter times. These parameter changes in the formulation allow the freeze-drying process to achieve high virus concentration without requiring extended drying periods, thus resolving the contradiction between concentration and time

Inventive Principle:
Principle #35Parameter changes

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 formulation achieves long-term stability and minimal virus titer loss, even at elevated temperatures, ensuring the biological activity of the vaccine is retained, making it suitable for industrial applications and global distribution.

Implementation Method 1

a freeze-dried formulation obtained by freeze-drying an aqueous formulation

Methodology Applied
Scientific EffectFreeze-drying: Freeze Drying

Implementation Method 2

Freeze-drying leads to dried biological products which are stable at about 4°C to 8 °C

Methodology Applied
Scientific EffectSublimation: Sublimation

Data Source

PatentEP2903600B1Virus-containing formulation and use thereof
Publication Date: 2022.08.31 TRANSGENE SA
  • EP2903600B1 patent drawingFigure 1A
  • EP2903600B1 patent drawingFigure 1B
  • EP2903600B1 patent drawing

AI summary

The present invention relates to formulation comprising (i) at least one virus-based material, (ii) at least one polymer selected in the group of polyvinylpyrrolidone and derivatives thereof, (iii) at least one sugar, (iv) at least two different amino acids, (v) at least two pharmaceutical acceptable salts, wherein at least one of said salts is a phosphate salt and, optionally (vi) a pharmaceutical acceptable buffer. Such a formulation is particularly suitable for freeze-drying. The present invention also relates to the corresponding dry product, as well as its preparation process. The present invention also relates to a reconstituted material comprising said dry product, which can be administered to a patient in need thereof. Such formulation and reconstituted material are useful as vaccines, preferably for the treatment and/or the prevention of cancers, infectious diseases and/or autoimmune disorders.