Multiplex Analyte Detection via Sample Segmentation

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current multiplex detection methods, such as proximity extension assays (PEA) and proximity ligation assays (PLA), face challenges in accurately detecting proteins with varying concentrations, as high-concentration proteins can overwhelm signals from low-concentration proteins, leading to failure in detecting the latter.

Innovation Solution

The method involves dividing a sample into multiple aliquots, each containing a subset of analytes selected based on predicted abundance, and performing separate multiplex assays for each aliquot, with the use of PCR reactions and internal controls to amplify and detect reporter nucleic acid molecules, ensuring accurate detection across a wide range of concentrations.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If multiplex detection is performed to detect multiple proteins in a single sample, then the ability to detect multiple analytes is improved, but the signal from high-concentration proteins overwhelms the signal from low-concentration proteins, resulting in failure to detect low-abundance analytes

Engineering Contradiction:

Improvemultiplex detection capabilityVSAvoiddetection accuracy for low-abundance analytes

Core Design Contradiction:

Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The sample is divided into multiple aliquots, and the analytes are segmented into different subsets based on their predicted abundance. Each aliquot is then processed separately to detect a specific subset of analytes, preventing signal overlap between high-abundance and low-abundance analytes while maintaining comprehensive multiplex detection capability

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

Different aliquots are assigned different detection sensitivities and analyte subsets tailored to their specific purposes. Aliquots detecting low-abundance analytes use optimized conditions for high sensitivity, while aliquots detecting high-abundance analytes use conditions optimized for their concentration range, allowing each local detection process to operate at optimal quality

Inventive Principle:

Principle #3Local quality

2Device complexity

If a single multiplex assay is used to detect all analytes, then the assay complexity is reduced, but the dynamic range of detection is limited due to signal saturation from high-concentration proteins

Engineering Contradiction:

Improveassay protocol simplicityVSAvoiddynamic range of detectable concentrations

Core Design Contradiction:

Device complexityVSQuantity of substance

Solution Approach 1:

The detection system is segmented into multiple parallel assays, each optimized for specific concentration ranges. This segmentation extends the overall dynamic range by ensuring that analytes across different concentration levels are detected in appropriate assays, while maintaining protocol simplicity through standardized processing steps

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The assay parameters such as probe concentrations, detection sensitivities, and signal thresholds are changed and optimized for different aliquots based on the predicted abundance of analytes in each aliquot, enabling detection across a wider dynamic range while maintaining manageable protocol complexity

Inventive Principle:

Principle #35Parameter changes

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 enables reliable detection of multiple analytes with varying concentrations, improving the accuracy of multiplex detection and allowing for precise quantification of biomarkers, particularly in the context of personalized medicine.

Implementation Method 1

nucleic acid moieties linked to the analyte-binding domains of a probe pair hybridise to one another when the probes are in close proximity

Methodology Applied

Scientific EffectNucleic acid hybridisation:

Implementation Method 2

extended using a nucleic acid polymerase. The extension product forms a reporter nucleic acid

Methodology Applied

Scientific EffectNucleic acid polymerase extension:

Implementation Method 3

a PCR reaction is performed to amplify the reporter nucleic acid molecule

Methodology Applied

Scientific EffectPCR amplification:

Implementation Method 4

a separate component which is present in a pre-determined amount, and which is, or comprises, or leads to the generation of, a control nucleic acid molecule which is amplified by the same primers as the reporter nucleic acid molecules

Methodology Applied

Scientific EffectPCR amplification:

Data Source

PatentUS20230159983A1Method for detecting analytes of varying abundance

Publication Date: 2023.05.25 OLINK PROTEOMICS AB

AI summary

The present invention provides a method of detecting multiple analytes in a sample, wherein said analytes have varying levels of abundance in the sample, said method comprising: (i) providing multiple aliquots from the sample; and (ii) in each aliquot, detecting a different subset of the analytes by performing a separate multiplex assay for each aliquot, wherein the analytes in each subset are selected based on their predicted abundance in the sample.