Single-Atom Nanozyme ELISA Biomarker Detection
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Solution Overview
Problem
ELISA techniques using HRP struggle to reliably detect biomarkers at low concentrations, such as those indicative of early-stage diseases like Alzheimer's, and are sensitive to environmental factors like pH and temperature, leading to short kit stability and limited detection capabilities.
Innovation Solution
The development of single-atom nano-enzymes (SANs) with metal moieties like iron, nitrogen, and carbon active sites, which are chemically linked to antibodies for enhanced catalytic capabilities and stability under harsh conditions, allowing for improved detection of low-concentration biomarkers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HRP is used as the enzyme label in ELISA, then the detection can be performed with standard procedures, but the detection sensitivity is insufficient for low concentration biomarkers and the kit stability is limited under environmental variations
Solution Approach 1:
The patent changes the fundamental parameter of the enzyme label from natural HRP to synthetic single-atom nano-enzymes (SANs) with metal moieties. This parameter change enables both enhanced catalytic activity for improved detection sensitivity and superior stability under varying environmental conditions, resolving the contradiction between measurement precision and reliability
Solution Approach 2:
The invention employs composite material structures where metal atoms (Fe, Co, Ni, Cu, or Mn) are anchored on nanoparticle supports (carbon dots, graphene quantum dots, or metal oxide nanoparticles). This composite approach creates SANs that combine the catalytic benefits of metal centers with the stability and surface area advantages of nanoparticle supports, achieving both high sensitivity and reliability
2Measurement precision
If HRP is used in ELISA kits, then the assay can detect biomarkers, but the detection limit is insufficient for early-stage disease diagnosis requiring sub-ten picogram per milliliter sensitivity
Solution Approach 1:
The patent fundamentally changes the catalytic parameter by replacing HRP with SANs that exhibit significantly higher catalytic activity. The metal-based SANs demonstrate enhanced turnover numbers and catalytic efficiency, enabling detection limits below 10 pg/mL necessary for early-stage disease diagnosis
Solution Approach 2:
The invention substitutes the biological enzyme system (HRP) with a synthetic chemical catalyst system (metal-based SANs). This substitution eliminates the limitations of natural enzymes while maintaining the desired catalytic function, achieving superior detection limits through enhanced catalytic capability
3Stability of the object's composition
If HRP-based ELISA kits are stored at low temperatures to maintain stability, then shelf life is extended, but the storage conditions become restrictive and operational flexibility is reduced
Solution Approach 1:
The patent changes the chemical composition parameter from organic protein-based HRP to inorganic metal-based SANs. This compositional change fundamentally improves thermal and pH stability, allowing the kits to maintain enzyme activity under varied storage conditions without requiring strict temperature control, thereby enhancing operational flexibility
4Productivity
If natural enzymes like HRP are used, then the catalytic activity is sufficient for standard applications, but the enzyme degrades quickly under harsh environmental conditions limiting kit shelf life
Solution Approach 1:
The invention uses composite material structures where metal atoms are anchored on stable nanoparticle supports (carbon dots, graphene quantum dots, or metal oxide nanoparticles). This composite architecture provides both high catalytic activity from the metal centers and long-term stability from the robust nanoparticle supports, achieving extended kit shelf life while maintaining productivity
Solution Approach 2:
The patent substitutes biodegradable natural enzymes with chemically stable synthetic metal-based catalysts. This substitution replaces the protein-based HRP system that degrades under harsh conditions with inorganic SANs that resist degradation, thereby extending kit shelf life while preserving catalytic activity
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
SANs demonstrate significantly increased catalytic performance and stability, enabling detection of biomarkers at lower concentrations with improved sensitivity and robustness against environmental variations, with a limit of detection an order of magnitude lower than traditional ELISA techniques.
Implementation Method 1
A SAN can include a nanoscale structure in which at least some or all the catalytic active sites contain a metal moiety (e.g., iron) present as isolated single atoms stabilized by the support of or by bonding with additional atoms
Implementation Method 2
Upon contact with the substrate, the HRP linked to the antibodies can catalyze an oxidation reaction between the TMB and hydrogen peroxide (H2O2) to produce a color change
Data Source
AI summary
Single atom nanozymes and associated immunoassays, method of making, and method of using such immunoassays are described herein. For example, a method of making a single atom nanozyme includes forming a soft template having multiple nanoscale structures in an aqueous solution and adding a monomer and a metal containing salt into the aqueous solution. The metal containing salt causes polymerization of the monomer to form multiple nanostructures according to the nanoscale structures of the soft template. The method also includes coating the individual formed nanostructures with a confinement layer in the aqueous solution before pyrolyzing. During pyrolysis, the confinement layer at least restricts or completely prevents migration of atoms on the external surface of the individual nanostructures.