Biomolecule Concentration via Segmented Diafiltration
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional ultrafiltration/diafiltration (UF/DF) processes struggle to produce highly concentrated biomolecule formulations with defined excipient contents, often resulting in residual buffer ions and impaired protein stability due to inadequate removal of anionic excipients, especially at high protein concentrations.
Innovation Solution
A modified UF/DF process involving an additional diafiltration step with a high ionic strength medium followed by diafiltration with water, reducing residual excipients to below detection levels and maintaining protein integrity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional UF/DF processes are used to concentrate biomolecules, then protein concentration is increased, but residual buffer ions and anionic excipients remain which impair protein stability
Solution Approach 1:
The diafiltration process is segmented into multiple sequential steps with different ionic strengths. The first diafiltration step uses a buffer with ionic strength similar to the starting material, while the second step uses water or low ionic strength buffer. This segmentation allows progressive removal of anionic excipients without causing protein precipitation or aggregation, resolving the contradiction between concentration and stability.
Solution Approach 2:
The ionic strength parameter of the diafiltration medium is systematically changed between steps. The process transitions from high ionic strength (first diafiltration) to low ionic strength (second diafiltration), enabling effective removal of residual excipients while maintaining protein stability throughout the concentration process.
2Manufacturing precision
If diafiltration is performed to remove excipients, then excipient content is reduced, but process time and complexity increase
Solution Approach 1:
The first diafiltration step performs preliminary removal of the majority of anionic excipients before the concentration step. This preliminary action reduces the excipient load early in the process, making the subsequent second diafiltration step more efficient and reducing the total number of diavolumes required to achieve the desired excipient content.
Solution Approach 2:
The process maintains continuous useful action by integrating diafiltration steps with concentration steps in a seamless sequence. The UF/DF operations continue without interruption, with the retentate from one step becoming the feed for the next, maximizing process efficiency while achieving precise excipient control.
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 process consistently achieves highly concentrated biomolecule formulations with reduced residual excipients, maintaining protein stability and allowing for easier purification, while minimizing physical stress on proteins.
Implementation Method 1
a first ultrafiltration UF1; a first diafiltration DF1; and a second diafiltration DF2; and a second ultrafiltration UF2
Data Source
AI summary
It is disclosed an improved multi-step process for the preparation of a highly concentrated liquid formulation containing biomolecules comprising the steps of (a) a first ultrafiltration UF1; (b) a first diafiltration DF1; (c) a second diafiltration DF2; and (d) a second ultrafiltration UF2; wherein an aqueous solution of one or more salts, as liquid medium B, is used for step (b) and water or an aqueous solution of one or more salts, as liquid medium C, is used for step (c), wherein the one or more salts used for step (b) are the same or different from the one or more salts used for step (c) and wherein the liquid medium B has an ionic strength which is higher than the ionic strength of the liquid medium C. The process according to the present invention allows the preparation of well-defined highly concentrated formulations containing biomolecules, particularly proteins, intended for pharmaceutical or non-pharmaceutical use. Unwanted excipient(s) of the starting liquid biomolecule formulation, may be reduced under solution conditions, to very low levels or levels lower than the detection limit.