Tangential Flow Filtration for High-Density Cell Culture Clarification
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Solution Overview
Problem
Current methods for purifying high cell density cell cultures are inefficient, often requiring dilution, expensive chromatography resins, and are prone to filter clogging, making them costly and unreliable for large-scale biopharmaceutical production.
Innovation Solution
The method involves processing crude cell culture harvests directly through tangential flow filtration (TFF) without preliminary dilution, using hollow fiber filters with specific pore sizes and diameters, to separate cells and proteins effectively, reducing the need for expensive resins and simplifying the process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high cell density culture is processed directly through filtration, then productivity is improved, but filter clogging occurs rapidly
Solution Approach 1:
The filtration process is divided into two distinct stages: first microfiltration to remove intact cells, then depth filtration to remove cell debris. This segmentation prevents clogging by handling different particle types in sequence rather than simultaneously, resolving the contradiction between processing speed and filter performance.
Solution Approach 2:
Microfiltration is performed as a preliminary step before depth filtration to remove intact cells. By eliminating the larger cell structures first, the subsequent depth filtration step can focus on smaller debris without becoming clogged, enabling direct processing of high cell density cultures without dilution.
2Reliability
If dilution is used to prevent filter clogging, then filter performance is maintained, but processing cost increases
Solution Approach 1:
The problematic intact cells are extracted and removed from the culture broth through microfiltration before the depth filtration step. This extraction eliminates the cause of clogging without requiring dilution, maintaining filter performance while avoiding the added complexity and cost of dilution steps.
3Manufacturing precision
If expensive chromatography resins are used for purification, then product purity is improved, but manufacturing cost increases
Solution Approach 1:
The invention replaces expensive, reusable chromatography resins with disposable depth filtration cartridges. Although the cartridges are single-use, their low cost and simplicity eliminate the need for expensive resin materials and complex regeneration procedures, achieving comparable purification at lower manufacturing cost.
Solution Approach 2:
The chemical adsorption mechanism of chromatography resins is replaced with a mechanical filtration system using depth filtration cartridges. This substitution maintains product purity through physical separation while eliminating the high costs associated with chromatography resin materials and operation.
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 approach allows for efficient clarification of high cell density suspensions with high protein recovery rates, reducing costs and increasing process reliability, enabling scalable and cost-effective purification of biopharmaceuticals like antibodies.
Implementation Method 1
processing the complete crude cell culture harvest obtained in step a) by tangential flow filtration (TFF), where the processed volume of the crude cell culture harvest is less than two times the volume of the bioreactor
Implementation Method 2
using hollow fiber filters with specific pore sizes and diameters, to separate cells and proteins effectively
Data Source
AI summary
The disclosure provides methods for the clarification of a cell broth with high cell density.