Surface Activation and Microarray Printing for Low-Background SPR
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Solution Overview
Problem
Existing surface plasmon resonance (SPR) assays face challenges in preventing non-specific binding of biomolecules, leading to high background signals and poor detection of target biomarkers due to non-specific interactions with assay surfaces, particularly in biofluids like serum, plasma, and whole blood.
Innovation Solution
A surface activation system (SAS) using thiolated protein A for functionalizing inert metal surfaces, combined with simultaneous printing of capture probes and blocking agents like modified PEG and serum proteins, to reduce non-specific binding and enhance detection sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If surface functionalization is performed to capture target biomarkers, then detection sensitivity is improved, but non-specific binding of biomolecules increases leading to high background signal
Solution Approach 1:
The patent applies different functional properties to different regions of the sensing surface. Capture probes are immobilized at specific locations to bind target biomarkers, while blocking agents are applied to surrounding areas to prevent non-specific binding. This spatial differentiation of functional properties allows simultaneous achievement of high detection sensitivity at probe locations and low background signal in non-probe areas.
Solution Approach 2:
Blocking agents serve as intermediary substances that mediate between the solid substrate and non-target biomolecules. These agents occupy non-specific binding sites on the surface, preventing unwanted proteins from adsorbing and forming non-specific interactions, thereby reducing background signal while allowing target-specific capture probes to function.
2Object-generated harmful factors
If blocking agents are applied to reduce non-specific binding, then background signal is reduced, but detection sensitivity may be compromised due to potential interference with target binding
Solution Approach 1:
The patent ensures that blocking agents are applied selectively to areas where capture probes are not present, or that the blocking agents used do not interfere with the specific binding interactions between capture probes and target biomarkers. This localized application or selective choice of blocking agents maintains low background signal while preserving detection sensitivity.
Solution Approach 2:
The patent optimizes parameters such as blocking agent concentration, incubation time, and blocking agent structure to achieve effective reduction of non-specific binding without compromising target detection. By carefully controlling these parameters, the system achieves optimal balance between low background signal and high detection sensitivity.
3Reliability
If surface functionalization is performed to improve biomarker detection, then assay specificity is improved, but assay complexity increases due to multiple treatment steps
Solution Approach 1:
The patent combines multiple functions into integrated surface treatment protocols. Surface functionalization and blocking steps are merged into a coordinated process where capture probes and blocking agents are applied in a sequence that achieves both specific target capture and non-specific binding prevention. This integration maintains high assay specificity while managing procedural complexity through systematic combination of steps.
Solution Approach 2:
The patent employs blocking agents and surface functionalization strategies that can be applied across different assay formats and target types. The surface treatment protocols are designed to be universally applicable to various biomarker detection scenarios, reducing the need for assay-specific customization and thereby managing complexity while maintaining reliability.
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 SAS effectively minimizes non-specific binding, improving the signal-to-noise ratio and enabling accurate detection of trace biomarkers in biofluids, enhancing the reliability of SPR-based assays.
Implementation Method 1
The surface agent typically is bifunctional, which can be immobilized on the surface, e.g., through a thiol group
Implementation Method 2
surface plasmon resonance (SPR) analysis
Data Source
AI summary
Provided herein are methods of surface functionalization, array construction, and/or blocking as well as substrates that are surface functionalized and blocked from non-specific bindings. Also provided herein are blocking solutions, blocking agents, and kits comprising the same. Further provided are methods of using functionalized and blocked surfaces for analyzing samples, such as in an SPR analysis.