Biomolecule Extraction Without Antibody Enrichment

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Solution Overview

Problem

Current methods for extracting and quantifying biomolecules from complex biological matrices are not selectively efficient, often resulting in contamination from other biomolecular classes, which can confound downstream detection and analysis, particularly for low-abundance proteins and peptides relevant to diseases like Alzheimer's.

Innovation Solution

A method using organic solvents for precipitation and liquid-liquid phase separation to extract biomolecules without antibody enrichment, followed by quantitation with stable isotope labeled standards for accurate concentration calculation, enabling detection and monitoring of biomarkers in biological fluids and tissues.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If antibody-based immunocapture is used to extract proteins/peptides, then selectivity for the target protein/peptide is improved, but the method becomes limited by antibody availability, species specificity, epitope accessibility, and cross-reactivity issues

Engineering Contradiction:

ImproveselectivityVSAvoidantibody dependency

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts the target protein/peptide from the complex biological matrix using organic solvent precipitation and liquid-liquid phase separation, removing the need for antibody-based capture. This extracts the essential function of selective enrichment while eliminating antibody dependency and its associated limitations.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent replaces the biochemical mechanism of antibody-antigen recognition with a physicochemical extraction mechanism using organic solvents. This substitution eliminates the biological complexity of antibody requirements while achieving selective extraction of target biomolecules based on their partitioning behavior between aqueous and organic phases.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

2Quantity of substance

If conventional extraction methods are used to enrich biomolecules, then broad biomolecular classes are captured, but contaminants from other biomolecular classes are co-enriched, confounding downstream detection

Engineering Contradiction:

Improvebiomolecule enrichmentVSAvoidextraction purity

Core Design Contradiction:

Quantity of substanceVSManufacturing precision

Solution Approach 1:

The patent applies local quality by creating distinct aqueous and organic phases with different solvation properties. Target proteins/peptides partition preferentially into one phase based on their local physicochemical characteristics, achieving selective enrichment while leaving contaminants in the other phase. This local differentiation of extraction properties enables high purity isolation.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the physical-chemical parameters of the extraction system by using organic solvents that alter the solubility and partitioning behavior of biomolecules. By adjusting solvent composition and phase separation conditions, the method achieves selective extraction of target biomolecules with high purity, resolving the contradiction between enrichment quantity and extraction purity.

Inventive Principle:

Principle #35Parameter changes

3Measurement precision

If antibodies are used for low-abundance protein/peptide capture, then detection sensitivity is improved, but the method is limited by optimization of incubation time and conditions

Engineering Contradiction:

Improvedetection sensitivityVSAvoidincubation time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent skips the lengthy antibody incubation and binding optimization steps by using rapid organic solvent-based extraction. The physicochemical partitioning process occurs quickly without requiring extended incubation times, achieving both high detection sensitivity for low-abundance proteins and rapid processing throughput.

Inventive Principle:

Principle #21Skipping (Rushing through)

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 allows for the precise extraction and quantification of low-abundance biomarkers, reducing contamination and enhancing the sensitivity and accuracy of biomolecule detection, suitable for diagnosing and monitoring diseases like Alzheimer's without the need for immunoprecipitation.

Implementation Method 1

A method using organic solvents for precipitation and liquid-liquid phase separation to extract biomolecules without antibody enrichment

Methodology Applied

Scientific EffectLiquid-liquid phase separation: Liquid-Liquid Extraction

Implementation Method 2

A method using organic solvents for precipitation and liquid-liquid phase separation to extract biomolecules

Methodology Applied

Scientific EffectPrecipitation: Precipitation

Data Source

PatentUS20180364260A1Methods for extracting and measuring concentrations of biomolecules in complex matrices without the need for immunocapture

Publication Date: 2018.12.20 C2N DIAGNOSTICS LLC

AI summary

Extraction and separation methods that can be used to quantify the absolute concentrations of one or more low abundance biomolecules present in biological fluids and tissues, without the need for antibody enrichment or reliance on an antibody for quantification are provided. These biomolecules can be biomarkers used to diagnose and monitor disease progression. As an example, these methods are applied to human plasma and CSF, and used to quantify Alzheimer's disease biomarkers.