Recapturing Proteins from Depth Filters Using High Salt Elution
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Solution Overview
Problem
Conventional methods for producing recombinant therapeutic proteins, such as Factor VIII, face challenges with low yields and high production costs due to protein binding with depth filters during purification, leading to reduced recovery and increased expenses.
Innovation Solution
The method involves treating used depth filters with a high salt elution buffer to recapture bound proteins of interest, utilizing a pseudo chromatographic approach to release and recover proteins from the filters, thereby increasing protein yield and efficiency in recombinant manufacturing processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If depth filtration is used to separate recombinant Factor VIII from source cells and debris, then purification is achieved, but protein recovery is reduced due to binding with the charged filter
Solution Approach 1:
The patent changes the ionic strength parameter of the buffer by introducing high salt concentration (e.g., 1M NaCl) to disrupt the electrostatic binding between the charged depth filter and the recombinant Factor VIII protein, thereby releasing the bound protein and improving recovery
Solution Approach 2:
The patent uses a high salt buffer as an intermediary substance that mediates the release of bound protein from the filter by competing for electrostatic interactions, allowing the protein to be recovered without direct mechanical disruption
2Productivity
If charged depth filters are used for separation, then purification efficiency is improved, but production cost increases due to reduced protein yield
Solution Approach 1:
By changing the ionic strength parameter through high salt buffer application, the patent reverses the adsorption equilibrium, releasing bound protein and increasing overall yield while maintaining the benefits of charged filter purification
Solution Approach 2:
The patent recovers the protein that would otherwise be discarded due to filter binding by applying elution conditions that release the bound protein, thereby converting a loss into a recoverable resource
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 enhances protein recovery and reduces production costs by effectively releasing bound proteins from depth filters, improving the overall yield and efficiency of recombinant protein production.
Implementation Method 1
exposing a filter through which the sample has been passed to an elution buffer wherein exposure to the elution buffer releases the protein of interest from the filter
Implementation Method 2
The charged depth filter binds to the FVIII product thereby reducing the final recovery of the protein
Data Source
AI summary
Embodiments disclosed herein provide systems and methods that increase protein yield from recombinant manufacturing processes. The systems and methods treat used depth filters with bound proteins of interest as a stationary phase exchange resin to recapture bound protein of interest from the depth filter.