Protein Purification via Binding Moieties Library
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Solution Overview
Problem
Current methods for protein purification, especially in biotechnology, face challenges in efficiently removing contaminating proteins like HCPs (Host Cell Proteins) from therapeutic protein solutions, as these proteins can cause severe allergic or immunological reactions, and existing techniques often require multiple steps reducing the overall yield of the target protein.
Innovation Solution
The use of non-selective libraries of binding moieties, such as combinatorial peptide libraries or antibody libraries, with a large number of diverse binding moieties to capture and separate contaminating proteins from the target protein, allowing for a single purification step that increases the purity of the target protein significantly.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If multiple purification steps are used to remove contaminating proteins, then the purity of the target protein is improved, but the overall yield and productivity deteriorate
Solution Approach 1:
The patent combines multiple purification functions into a single affinity chromatography step using a fusion protein that simultaneously captures the target protein and removes contaminating proteins. This merging of functions eliminates the need for sequential purification steps, thereby maintaining high purity while preserving overall yield and productivity.
Solution Approach 2:
The affinity ligand in the patent serves multiple functions: it specifically binds the target protein for capture, and simultaneously enables removal of contaminating proteins through the fused purification domain. This multi-functionality allows a single reagent to perform what traditionally required multiple separate purification steps.
2Manufacturing precision
If affinity chromatography is used to purify the target protein, then the purity is improved, but the cost and device complexity increase
Solution Approach 1:
The patent merges the affinity recognition domain with a protein purification domain into a single fusion protein. This combination eliminates the need for separate affinity ligand preparation and purification steps, simplifying the overall process while maintaining the high purity benefits of affinity chromatography.
3Measurement precision
If antibodies are used for immunoaffinity chromatography to remove specific contaminants, then the specificity is improved, but the cost increases and the method is not feasible when contaminant identity is unknown
Solution Approach 1:
The patent segments the purification function into two independent domains within a fusion protein: the affinity recognition domain that provides specificity for the target protein, and the purification domain that removes contaminants. This segmentation allows the method to achieve specific contaminant removal without requiring prior knowledge of contaminant identity, as the purification domain acts on a broad range of proteins.
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 effectively removes a high percentage of contaminating proteins from the target protein solution in a single step, enhancing the purity of the target protein to at least 95% and maintaining a significant yield, thereby reducing the risk of adverse reactions and improving the efficiency of the purification process.
Implementation Method 1
contacting a sample comprising at least 95% of the target protein group and at most 5% of contaminating proteins with a library having at least 100 different binding moieties in an amount sufficient to bind contaminating proteins and a minority of the target protein group
Data Source
AI summary
The present invention provides methods and kits for purifying a target protein group. The method comprises the steps of contacting a sample comprising at least 95% of the target protein group and at most 5% of contaminating proteins with a library of binding moieties having different binding moieties, binding the contaminating proteins and a minority of the target protein group to the library of binding moieties, separating the unbound target protein group from the proteins bound to the library of binding moieties and collecting the unbound target protein. The collected target protein is more pure than the target protein group in the sample.