Separating Agent Lysine Substitution Dynamic Binding Capacity
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Solution Overview
Problem
Binding a single-chain antibody to a carrier via its lysine residue can reduce the dynamic binding capacity (DBC) of the separating agent for target substances due to the amino acid sequence, limiting its effectiveness in separating target substances from mixed liquids.
Innovation Solution
Substituting lysine residues in the single-chain antibody with other amino acid residues and incorporating an amino acid sequence with three or more lysine residues at the C-terminus, which are then chemically bonded to a carrier surface, enhancing the DBC of the separating agent.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the single-chain antibody is bound to the carrier via its original lysine residue, then the separating agent can be produced, but the dynamic binding capacity (DBC) to the target substance is reduced
Solution Approach 1:
The antibody molecule is segmented functionally: the variable region (which binds the target substance) is separated from the constant region (which contains the added lysine residues for carrier binding). This segmentation allows the binding site to remain intact while providing dedicated attachment points elsewhere on the molecule, thus resolving the contradiction between maintaining binding capacity and achieving high productivity through efficient carrier immobilization.
Solution Approach 2:
Additional lysine residues are introduced as intermediary elements at the C-terminus of the constant region. These intermediary lysine residues serve as dedicated connection points between the antibody and the carrier, mediating the binding process without interfering with the original antigen-binding function of the variable region. This allows efficient carrier binding while preserving dynamic binding capacity.
2Productivity
If lysine residues in the variable region are substituted with other amino acid residues, then the dynamic binding capacity is improved, but the binding efficiency may be affected
Solution Approach 1:
Amino acid substitutions are applied locally and selectively only to lysine residues in the variable region, while the rest of the antibody structure remains unchanged. This localized modification approach improves dynamic binding capacity by preventing unwanted interactions in the variable region, while preserving the overall binding efficiency through maintenance of the antibody's core structure and function.
Solution Approach 2:
The chemical properties of amino acid residues in the variable region are changed by substituting lysine residues with other amino acids. This parameter change (altering charge distribution and chemical reactivity) prevents unwanted interactions that would reduce dynamic binding capacity, while the substitutions are carefully selected to maintain binding efficiency to the target substance.
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 significantly improves the dynamic binding capacity of the separating agent, enabling more effective separation of target substances from mixed liquids, as demonstrated by increased protein immobilization and capture efficiency.
Implementation Method 1
a surface of the carrier and lysine residues in the amino acid sequence containing three or more lysine residues are bound by chemical bonds
Data Source
AI summary
Provided is a separating agent with an improved dynamic binding capacity (DBC) to a target substance. The separating agent includes a carrier and a protein, wherein the protein is a given protein, and a surface of the carrier and added lysine residues in the protein are bound by chemical bonds.