Oriented Protein Ligand Immobilization on Chromatography Support
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Solution Overview
Problem
Conventional affinity chromatography methods face inefficiencies in immobilizing protein ligands due to disorderly binding, which reduces the ligand's binding capacity and utilization in immunoglobulin purification.
Innovation Solution
An affinity support is developed with a linker containing an amino acid residue with an amino or thiol group for oriented binding to a solid phase support, incorporating an immunoglobulin-binding domain from Fc-binding proteins or Protein A, and using an epoxy group for covalent binding to maintain ligand orientation and enhance binding capacity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If the ligand is chemically bonded to various functional groups on the support surface, then the ligand can be immobilized on the support, but the multiple functional groups are disorderly bound causing insufficient effectiveness
Solution Approach 1:
The patent introduces a linker molecule as an intermediary between the support surface and the ligand. The linker contains a specific functional group that reacts with the support surface, while the other end is designed to bind to the ligand in a controlled manner, preventing disorderly binding and maintaining ligand effectiveness
Solution Approach 2:
The patent applies local quality by creating a specific binding region on the support surface through the linker. Instead of allowing random bonding across the entire surface, the linker creates localized, oriented binding sites that ensure proper ligand orientation and maintain binding capacity
2Productivity
If conventional affinity chromatography is used, then the ligand can be immobilized on the support, but the binding capacity is reduced due to disorderly orientation
Solution Approach 1:
The patent applies preliminary action by pre-designing the linker structure with specific functional groups positioned to ensure proper ligand orientation before the actual immobilization process. This pre-arranged structure guides the ligand into the correct orientation during binding, eliminating disorderly arrangement
Solution Approach 2:
The patent changes the chemical parameters of the immobilization process by using a linker with specific functional groups (amino or thiol groups) that react with epoxy groups on the support surface. This chemical parameter change enables controlled, oriented binding rather than random attachment
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 affinity support efficiently immobilizes protein ligands with improved binding properties, achieving high immunoglobulin purification efficiency and cost-effectiveness by preventing ligand misorientation and maintaining high binding capacity.
Implementation Method 1
the ligand is chemically bonded to various functional groups existing on a surface of the support through functional groups on the ligand
Implementation Method 2
An affinity chromatography is a chromatography using a column filled with a ligand immobilization support in which a substance (ligand) capable of specifically binding a substance for the purpose of separation or purification
Data Source
AI summary
To provide an affinity support in which a binding property of a ligand to a target substance is improved. The affinity support contains a solid phase support and a protein ligand, wherein the protein ligand is represented by formula (1): R-R1 (1) wherein R represents a linker binding to the solid phase support, which contains a polyproline, and R1 represents a protein showing an affinity to immunoglobulin, and the R is bound to a C terminal or an N terminal of an amino acid sequence in R1.