Monophasic Solvent and Bead-Based Multi-Omic Sample Preparation
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Solution Overview
Problem
Current multi-omic sample preparation methods for mass spectrometry are laborious, disparate, and difficult to automate, requiring numerous pipetting, vortexing, and centrifugation steps, and often take 1-2 days to complete, leading to low throughput and excessive sample loss.
Innovation Solution
A method involving a monophasic extraction solvent and paramagnetic bead technology for on-bead protein aggregation, allowing for rapid protein digestion and simplifying the extraction of lipids, metabolites, and proteins from a single sample, reducing preparation time to approximately three hours.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If conventional multi-omic sample preparation methods are used, then comprehensive extraction of lipids, metabolites, and proteins is achieved, but the process requires numerous pipetting, vortexing, and centrifugation steps taking 1-2 days to complete
Solution Approach 1:
The patent combines extraction of lipids, metabolites, and proteins into a single simultaneous process using a monophasic solvent system, eliminating the need for separate extraction steps. The monophasic solvent allows all three biomolecule classes to be extracted together in one procedure, reducing the number of processing steps from multiple sequential operations to a single integrated operation.
Solution Approach 2:
The monophasic solvent system serves multiple functions simultaneously: it extracts lipids, metabolites, and proteins from the sample in a single operation. This universal solvent system replaces multiple specialized extraction procedures, enabling comprehensive multi-omic analysis through one unified extraction step.
2Productivity
If conventional multi-omic sample preparation methods are used, then biomolecule extraction is achieved, but throughput is low and sample loss is excessive
Solution Approach 1:
By merging all three extraction operations into a single simultaneous procedure, the method increases throughput because multiple samples can be processed in parallel during the same time period. The unified extraction approach eliminates sequential bottlenecks and reduces the total processing time per sample, thereby increasing overall laboratory throughput.
Solution Approach 2:
The monophasic solvent system performs self-optimization by automatically distributing and extracting all three biomolecule classes in one operation without requiring multiple specialized steps. This self-sufficient extraction mechanism reduces manual intervention and minimizes sample loss that occurs during multiple transfer and processing steps.
3Extent of automation
If conventional multi-omic sample preparation methods are used, then comprehensive analysis is achieved, but the workflow is difficult to automate
Solution Approach 1:
The consolidation of three separate extraction workflows into a single unified procedure using monophasic solvent creates an automated-friendly workflow. With fewer discrete steps requiring manual intervention, the process can be more easily integrated with robotic automation systems and high-throughput platforms, improving both automation extent and operational ease.
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 streamlines sample preparation, reducing the number of steps and time by 94%, making it more amenable to robotic automation and multi-well plate formats, while maintaining comparable biomolecule coverage and data quality to standard workflows.
Implementation Method 1
The extraction solvent is able to solubilize a first portion of biomolecules, which optionally comprises lipids, carbohydrates, metabolites, and combinations thereof
Implementation Method 2
The plurality of immobilizing beads are able to bind and immobilize a second portion of biomolecules, which optionally comprises nucleic acids, proteins, polypeptides, and combinations thereof
Data Source
AI summary
Multi-omic analysis (analysis of proteins, lipids, and metabolites) is a powerful and increasingly utilized approach to gain insight into complex biological systems. One major hindrance with multi-omics, however, is the lengthy sample preparation process. Preparing samples for mass spectrometry (MS)-based multi-omics broadly involves extraction of metabolites and lipids with organic solvents, precipitation of proteins, and overnight digestion of proteins. The existing workflows are disparate and laborious, requiring multiple complex operation steps typically taking 1-2 days to perform. The present invention provides methods for preparing multi-omic samples that are faster and simpler than conventional methods, making it easier for a single lab or researcher to collect quality multi-omic data. A monophasic extraction solvent is used to efficiently extract biomolecules from a sample, including lipids and both polar and non-polar metabolites, and is paired with on-bead protein aggregation and rapid protein digestion.