Systems and methods for sample preparation, data generation, and protein corona analysis
Inventors
Manning, William • Kim, Young • Kwan-Leong, Brandon • Liou, Hope • Zhao, Xiaoyan • Hornburg, Daniel • Goldberg, Martin
Assignees
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Publication Number
US-12345715-B2
Publication Date
2025-07-01
Expiration Date
Abstract
Systems and methods for automated sample preparation and processing of protein corona are described herein, as well as its application in the discovery of advanced diagnostic tools as well as therapeutic agents.
Core Innovation
The invention provides an automated system for preparing a plasma or serum sample for mass spectrometry analysis using a plurality of magnetic particles. The system includes a fluid transfer unit, a magnetic unit comprising a magnet, partitions each comprising the plurality of magnetic particles, and a control unit comprising one or more processors with a program to perform steps for binding biomolecules and preparing material for mass spectrometry. The magnetic particles comprise an ammonium group and a positive surface charge and are configured to be immobilized when the magnetic unit is placed in proximity to the magnetic particles.
The system incubates the plasma or serum sample with the plurality of magnetic particles so that biomolecules comprising lipids and at least 300 different proteins bind to the magnetic particles. The bound particles are immobilized, the biomolecules bound to the particles are isolated from a supernatant, and the bound biomolecules are washed. The washed biomolecules are then contacted with a reducing agent configured to reduce the at least 300 different proteins.
The system prepares a population of peptides for mass spectrometry by incubating a digestion solution with the washed biomolecules and the plurality of magnetic particles. The digestion solution comprises a lysing reagent, producing peptides suitable for mass spectrometry analysis.
Claims Coverage
The independent claim covers an automated plasma or serum-to-peptide mass spectrometry sample preparation workflow using magnetic particles with defined surface chemistry and a reduced-protein digestion pathway, including a control unit that executes the preparation steps.
Automated plasma or serum mass spectrometry preparation using immobilizable ammonium positively charged magnetic particles
An automated system including a fluid transfer unit, a magnetic unit with a magnet, and partitions containing magnetic particles that comprise an ammonium group and a positive surface charge and are configured to be immobilized when the magnetic unit is placed in proximity.
Binding lipids and at least 300 proteins to magnetic particles from plasma or serum
A control unit executing incubating the plasma or serum sample with the plurality of magnetic particles under conditions sufficient to bind biomolecules comprising lipids and at least 300 different proteins to the plurality of magnetic particles.
Reducing bound proteins and producing peptides using a digestion solution with a lysing reagent
Steps to immobilize the magnetic particles, isolate biomolecules bound to the magnetic particles from a supernatant, wash the bound biomolecules, contact the washed biomolecules with a reducing agent configured to reduce the at least 300 different proteins, and prepare a population of peptides for mass spectrometry by incubating a digestion solution with the washed biomolecules and the magnetic particles, wherein the digestion solution comprises a lysing reagent.
Overall claim coverage centers on automated sample preparation for mass spectrometry by binding lipids and a large protein set to immobilizable ammonium/positively charged magnetic particles, then isolating, washing, reducing bound proteins, and generating peptides via digestion with a lysing reagent.
Stated Advantages
Enables binding biomolecules comprising lipids and at least 300 different proteins to the magnetic particles.
Supports preparing a population of peptides for mass spectrometry after reduction and digestion using a lysing reagent.
Documented Applications
Preparing a plasma or serum sample for mass spectrometry analysis.
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