Repebody IgG Purification via Neutral pH Elution
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Solution Overview
Problem
Current methods for purifying immunoglobulin G (IgG) using protein A are costly and damage antibodies due to acidic elution conditions, and the stability of protein A in high-pH solutions limits column reuse, increasing production costs.
Innovation Solution
Development of a repebody, a polypeptide selectively bound to IgG, which is produced using a recombinant microorganism and fused with a Leucine rich repeat (LRR) family protein and variable lymphocyte receptor (VLR) protein, allowing for immobilization and purification of IgG under milder conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If protein A is used for IgG purification, then purification effectiveness is improved, but production cost increases and antibody damage occurs due to acidic elution conditions
Solution Approach 1:
The patent employs a repebody column that can be reused multiple times without degradation, replacing the disposable nature of protein A columns. The repebody maintains stability across multiple purification cycles, eliminating the need for frequent column replacement and reducing overall production costs while avoiding antibody damage through neutral pH elution
Solution Approach 2:
The invention changes the elution pH parameter from acidic (pH 2-3) to neutral pH conditions. This parameter change eliminates antibody damage while maintaining purification effectiveness, as the repebody retains binding capability at neutral pH unlike protein A which requires acidic conditions for elution
2Productivity
If protein A column is reused in high-pH solution, then production process continues, but column stability decreases and reuse times are reduced
Solution Approach 1:
The repebody is engineered to be stable in high-pH solutions, allowing the column to be regenerated and reused under alkaline conditions without degradation. This parameter change in stability profile enables continuous production while maintaining column integrity across multiple reuse cycles
3Ease of operation
If acidic elution conditions are used for protein A, then IgG elution is achieved, but antibody structure is damaged
Solution Approach 1:
The invention changes the elution pH from acidic to neutral conditions. The repebody maintains its binding and elution functionality at neutral pH, eliminating the structural damage caused by acidic conditions while preserving ease of operation and elution capability
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 repebody enables efficient and cost-effective purification of IgG with improved stability and increased reuse of purification columns, reducing the risk of antibody damage and lowering production costs.
Implementation Method 1
a repebody selectively bound to an immunoglobulin G... having a binding capacity against immunoglobulin G... selectively bound to the immunoglobulin G
Data Source
AI summary
The present invention relates to a polypeptide (repebody) selectively bound to an immunoglobulin G, a polynucleotide which encodes the repebody, a vector containing the polynucleotide, a recombinant microorganism in which the polynucleotide is introduced, a method for producing the repebody using the recombinant microorganism, and a method for immobilizing or purifying an immunoglobulin G using the repebody. The repebody according to the present invention is useful as utilized for immobilization of an immunoglobulin G, purification of an immunoglobulin G, and production of an immunosensor, since the repebody selectively bound to an immunoglobulin G.