Multiplexed Assay Separating Binding and Detection Steps
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Solution Overview
Problem
Current multiplexed assays for detecting analytes in samples are time-consuming due to the integration of binding and detection steps, which limits their efficiency and speed.
Innovation Solution
A method is described where the analyte binding step is separated from the detection step, involving the formation of a binding complex with a detection reagent that includes an analyte binding portion, a targeting reagent, and a label, followed by a release step to isolate a detectable portion, which is then transferred to an assay surface for detection, allowing for faster analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the analyte binding step and detection step are integrated in current multiplexed assays, then the assay can detect analytes with sufficient sensitivity, but the overall assay time is prolonged and efficiency is reduced
Solution Approach 1:
The assay is divided into distinct binding step and detection step. During the binding step, capture molecules bound to particles capture analytes from the sample. During the detection step, detection molecules with labels are added to form sandwich complexes. This segmentation allows optimization of each step independently and enables faster overall assay performance while maintaining detection sensitivity.
Solution Approach 2:
Capture molecules are pre-immobilized on particles before the assay begins. During the binding step, these pre-positioned capture molecules immediately capture analytes from the sample without requiring additional preparation time. This preliminary positioning of binding components accelerates the overall assay process while ensuring sensitive detection.
2Productivity
If the analyte binding step and detection step are separated into distinct steps, then the overall assay time is reduced and efficiency is improved, but the procedural complexity increases
Solution Approach 1:
Particles serve multiple functions: they provide a solid support for immobilizing capture molecules, enable magnetic separation for washing away unbound components, and facilitate concentration of analytes. This multi-functionality reduces the need for additional specialized components, simplifying the overall procedure despite the separated binding and detection steps.
Solution Approach 2:
Sandwich complexes act as intermediaries that bridge the binding step and detection step. The sandwich complex consists of a capture molecule bound to the particle, the captured analyte, and a detection molecule with a label. This intermediary structure allows efficient transfer of the analyte from the binding phase to the detection phase, streamlining the separated steps.
3Adaptability or versatility
If sandwich complexes are formed with particulate support surfaces for multiplexed detection, then multiple analytes can be detected simultaneously, but the complexity of forming and managing multiple particle types increases
Solution Approach 1:
Different particle types are used with distinct magnetic properties (e.g., different coercivity values) that allow selective manipulation. Each particle type is functionally specialized for capturing specific analytes, enabling multiplexed detection while simplifying particle management through property-based differentiation rather than complex procedural handling.
Solution Approach 2:
Particles are differentiated by physical parameters such as magnetic coercivity, size, or surface properties. These parameter changes enable selective separation and detection of different analyte-particle complexes using magnetic field strength variations, simplifying the management of multiple particle types in multiplexed assays.
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 reduces the overall time required for the assay, enabling faster and more efficient detection of analytes by separating the binding and detection processes, thereby improving the speed and efficiency of multiplexed assays.
Implementation Method 1
the presence of a target analyte is indicated by the presence or absence of an observable label attached to one or more binding materials
Implementation Method 2
the specificity of many biochemical and biological binding reactions using binding partners such as antigen-antibody, complementary nucleic acids, or protein-ligand binding partners
Implementation Method 3
electrochemiluminescence can be triggered by a voltage imposed on a working electrode at a particular time and in a particular manner. The light produced by the label is measured and indicates the presence or quantity of the analyte
Implementation Method 4
magnetic or paramagnetic particles
Data Source
AI summary
Described herein are methods, systems and kits for conducting an assay for one or more analytes of interest in a sample. In one aspect, a method is provided for conducting a multiplexed binding assay for a plurality of analytes of interest in a sample. In one aspect, the assay includes an analyte binding step and a detection step. In one aspect, the analyte binding step is separated from the detection step.