Fluidized Bed Centrifuge Separation for Aqueous Two-Phase Bioprocessing

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Solution Overview

Problem

Existing methods for separating viral vectors from cell broth in bioreactors face challenges due to the small size of viral vectors, which are similar in size to cell debris, making efficient extraction difficult, and centrifugal extractors require frequent adjustments when phase volumes change.

Innovation Solution

Utilizing a fluidized bed centrifuge to separate the phases of an aqueous-two-phase system, where the centrifuge is loaded with the system as a fluid flow, allowing phases to separate based on density differences, with adjustments made via chamber filling volume and monitored by conductivity sensors.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a centrifugal extractor is used to separate phases of an aqueous-two-phase system, then continuous separation is achieved, but frequent adjustment of the weir is required when relative phase volumes change

Engineering Contradiction:
Improvecontinuous separationVSAvoidweir adjustment frequency
Core Design Contradiction:
ProductivityVSEase of operation

Solution Approach 1:

The patent employs a fluidized bed centrifuge where the separation mechanism dynamically adapts to changing phase volumes. The fluidized bed structure allows the separation medium to be continuously circulated and adjusted, enabling the system to automatically compensate for volume changes without manual weir adjustment. The fluid flow rates and bed characteristics can be dynamically modified to maintain optimal separation conditions.

Inventive Principle:
Principle #15Dynamics

Solution Approach 2:

The patent changes operational parameters including fluid flow rates, centrifugal force, and bed characteristics to adapt to varying phase volumes. By adjusting these parameters, the system maintains efficient separation across different operating conditions without requiring physical reconfiguration of the separation device.

Inventive Principle:
Principle #35Parameter changes

2Ease of operation

If fluidized bed centrifugation is used to separate small particles like cell debris and bacteria, then mild separation of larger cells is achieved, but small particles below 5 μm cannot be effectively separated

Engineering Contradiction:
Improvemild separation conditionVSAvoidseparation of small particles
Core Design Contradiction:
Ease of operationVSManufacturing precision

Solution Approach 1:

The patent segments the separation process into multiple stages: first separating larger cells gently, then using the fluidized bed to separate smaller particles through controlled fluid flow and centrifugal forces. This multi-stage approach allows different particle sizes to be separated effectively under appropriate conditions for each stage.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent utilizes hydraulic principles by circulating fluid through the fluidized bed at controlled rates. The fluid flow creates separation forces that can be adjusted to target specific particle sizes. By optimizing flow rate, viscosity, and pressure gradients, the system achieves separation of small particles below 5 μm while maintaining the mild conditions needed for cell integrity.

Inventive Principle:
Principle #29Pneumatics and hydraulics

3Manufacturing precision

If aqueous-two-phase systems are used to extract viral vectors, then separation based on density is achieved, but the small size of viral vectors (below 5 μm) makes them indistinguishable from cell debris

Engineering Contradiction:
Improvedensity-based separationVSAvoidsize discrimination capability
Core Design Contradiction:
Manufacturing precisionVSDifficulty of detecting and measuring

Solution Approach 1:

The patent introduces an intermediary substance - the fluidized bed separation medium - that provides an additional separation mechanism. This intermediary creates a controlled environment where density differences can be amplified and detected, allowing viral vectors to be separated from cell debris through the fluidized bed's selective retention and release characteristics.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces direct mechanical size-based separation with a fluid dynamics-based separation system. Instead of relying on physical size differences that are difficult to detect, the system uses fluid flow, centrifugal forces, and density gradients to achieve separation. This substitution enables detection and measurement of viral vectors based on their density and interaction with the fluidized bed rather than direct size measurement.

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

Enables efficient separation of viral vectors from cell debris with minimal operational complexity, allowing for selective purification and high yield of viral vectors while managing varying phase volumes.

Implementation Method 1

the aqueous-two-phase system is loaded into a rotating chamber of the fluidized bed centrifuge as a fluid flow, that due to the rotation of the chamber the aqueous-two-phase system is centrifuged such that a first phase of the two phases is separated from a second phase of the two phases

Methodology Applied
Scientific EffectCentrifugal separation: Centrifugal Separation

Implementation Method 2

The phases are separated due to a density difference between the phases, as is generally the principle of centrifugation

Methodology Applied
Scientific EffectDensity difference: Density Gradient

Implementation Method 3

an electrical conductivity can be measured to detect the different phases

Methodology Applied
Scientific EffectElectrical conductivity measurement: Conduction (electrical)

Data Source

PatentEP4632058A1Method of operating a process arrangement, in particular bioprocess arrangement
Publication Date: 2025.10.15 SARTORIUS STEDIM BIOTECH GMBH
  • EP4632058A1 patent drawingFigure 1
  • EP4632058A1 patent drawingFigure 2
  • EP4632058A1 patent drawingFigure 3

AI summary

The invention relates to a Method of operating a process arrangement (1), in particular bioprocess arrangement, wherein the method comprises a separation of two phases of an aqueous-two-phase system (2). It is proposed that the process arrangement (1) comprises a fluidized bed centrifuge (3), that the fluidized bed centrifuge (3) is used for the separation of the two phases of the aqueous-two-phase system (2), that the aqueous-two-phase system (2) is loaded into a rotating chamber (4) of the fluidized bed centrifuge (3) as a fluid flow, that due to the rotation of the chamber (4) the aqueous-two-phase system (2) is centrifuged such that a first phase (7) of the two phases is separated from a second phase (8) of the two phases, and, that due to the fluid flow during the rotation of the chamber (4), the first phase (7) remains in the chamber (4) while the second phase (8) leaves the chamber (4), such that the two phases are separated.