Mutant Immunoglobulin-Binding Domain for Higher Affinity Support Capacity
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Solution Overview
Problem
Existing affinity supports for immunoglobulin purification face challenges due to reduced immunoglobulin-binding capacity, leading to increased costs and yield loss, despite improvements suggested by previous patent literatures.
Innovation Solution
An affinity support using a mutant immunoglobulin-binding domain with specific amino acid mutations, such as substitutions or deletions at positions 58, 50, 49, and 42, and the inclusion of Lys at positions 4, 7, and 35, enhances the immunoglobulin-binding capacity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the immunoglobulin-binding domain is immobilized onto the support via chemical bonding using multiple functional groups, then the domain can be stably attached to the support surface, but the domain bonds in a disordered orientation which causes steric hindrance and reduces immunoglobulin-binding capacity
Solution Approach 1:
The patent extracts the problematic C-terminal lysine residue that causes disordered bonding, and separately removes or modifies it to enable controlled orientation. The immunoglobulin-binding domain is taken out from its natural state and re-engineered with specific mutations to achieve proper orientation while maintaining stable attachment.
Solution Approach 2:
The patent applies local quality by making specific mutations at particular positions (C-terminal lysine deletion or substitution, and mutations at positions 4, 7, 35, 42, 49, 50, 58) while keeping the rest of the domain structure intact. This localized modification approach enables controlled orientation without affecting the overall binding functionality.
2Productivity
If the immunoglobulin-binding capacity is improved through mutant domains, then the yield and cost-efficiency increase, but the domain structure and bonding characteristics must be precisely controlled
Solution Approach 1:
The patent changes the chemical parameters of the domain by introducing specific amino acid mutations (substitutions at positions 4, 7, 35, 42, 49, 50, 58 and C-terminal lysine modification). These parameter changes in the amino acid sequence directly control the bonding orientation and improve immunoglobulin-binding capacity, leading to higher purification yield.
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 mutant immunoglobulin-binding domain improves the immunoglobulin-binding capacity, resulting in higher yield and reduced costs for immunoglobulin purification.
Implementation Method 1
Protein A has five immunoglobulin-binding domains called E domain, D domain, A domain, B domain, and C domain, and each domain, by itself, can bind to an immunoglobulin
Data Source
AI summary
An affinity support having improved binding capacity for target proteins. An immunoglobulin-binding protein including a mutant immunoglobulin-binding domain, an affinity support including a solid-phase support and the immunoglobulin-binding protein immobilized thereto. The mutant immunoglobulin-binding domain consists of an amino acid sequence having at least 85% identity with an amino acid sequence of any of SEQ ID NOs: 1 to 12.