Silane Coupling Agent Immobilization for Protein Density
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Solution Overview
Problem
Current methods for immobilizing proteins on solid phase carriers suffer from high non-specific adsorption, leading to insufficient protein density and poor signal-to-noise ratios in biochemical assays, despite using porous structures and blocking agents.
Innovation Solution
An immobilization method involving contact of a solid phase carrier with an acid anhydride functional group-containing silane coupling agent, followed by binding of the protein at temperatures between 0°C and 60°C, without baking, to reduce non-specific adsorption and enhance protein density.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If a silane coupling agent is used to immobilize protein on a solid phase carrier, then the protein can be immobilized at a relatively high density, but non-specific adsorption of the substrate increases
Solution Approach 1:
The patent changes the temperature parameter from conventional high temperature (≥100°C) to a lower temperature range (0-60°C), which fundamentally alters the immobilization mechanism and reduces non-specific adsorption while maintaining protein density
Solution Approach 2:
The patent introduces a blocking agent as an intermediary substance that occupies non-specific binding sites on the carrier surface, preventing substrate adsorption while allowing specific protein immobilization to proceed
2Quantity of substance
If a porous structure is used to increase the amount of immobilization, then the amount of ligand can be increased, but non-specific adsorption also increases
Solution Approach 1:
The patent changes the temperature parameter to lower ranges (0-60°C), which suppresses non-specific adsorption mechanisms that are typically enhanced in porous structures at high temperatures, while still allowing sufficient ligand immobilization
Solution Approach 2:
The blocking agent serves as a mediator that selectively blocks non-specific binding sites within the porous structure, allowing the porous structure to maintain its high surface area and ligand capacity while preventing harmful non-specific adsorption
3Object-affected harmful factors
If a blocking agent is used to suppress non-specific adsorption, then non-specific adsorption is suppressed, but the S/N ratio does not increase
Solution Approach 1:
The patent changes the temperature parameter to lower ranges, which simultaneously achieves both non-specific adsorption suppression and enhanced S/N ratio by optimizing the balance between blocking agent effectiveness and signal generation
Solution Approach 2:
The patent uses a controlled amount of blocking agent that is sufficient to suppress non-specific adsorption but not excessive to the point of blocking all binding sites, thereby maintaining an optimal S/N ratio
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 method achieves significant reduction in non-specific adsorption and enhances the signal-to-noise ratio by allowing a higher density of protein immobilization on the carrier surface, improving the specificity and sensitivity of biochemical assays.
Implementation Method 1
bringing the solid phase carrier into contact with an acid anhydride functional group-containing silane coupling agent
Implementation Method 2
carrying out a process of binding of the physiologically active substance to the acid anhydride functional group while maintaining the solid phase carrier after the contact at a temperature within the range of 0° C. to 60° C.
Data Source
AI summary
An immobilization method for immobilizing a physiologically active substance on a solid phase carrier, the method including: bringing the solid phase carrier into contact with an acid anhydride functional group-containing silane coupling agent represented by the following Formula (I); and carrying out a process of binding of the physiologically active substance to the acid anhydride functional group while maintaining the solid phase carrier after the contact at a temperature within the range of 0° C. to 60° C.; a physiologically active substance-immobilized carrier, and a carrier for immobilization are provided. Further, a carrier including a porous material treated with an acid anhydride functional group-containing silane coupling agent represented by the following Formula (I), a blocking agent that is immobilized to the porous material; and a method for producing it is provided.