Blood preparation and profiling
Inventors
Karsten, Elisabeth • Herbert, Ben • Liddle, Alan • Hill, Cameron
Assignees
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Publication Number
US-11243218-B2
Publication Date
2022-02-08
Expiration Date
Abstract
The present disclosure relates to methods for generating blood protein profiles in red blood cell-enriched blood samples. The disclosed methods represent a new and improved laboratory technique for producing a protein profile from blood, increasing protein detection.
Core Innovation
The invention relates to producing a protein profile by obtaining a blood sample, separating plasma from the blood sample to produce a cell pellet, and leukodepleting the cell pellet to produce a red blood cell-enriched sample. The method incubates a small volume of the red blood cell-enriched sample in a medium, and one or more proteins secreted or released from the red blood cell-enriched sample into the incubated medium are detected, so the produced protein profile comprises the detected one or more proteins in the incubated medium.
In a related approach, the invention produces a protein profile by measuring the level of one or more proteins in the red blood cell-enriched sample and measuring the level of the same one or more proteins in the separated plasma. The method calculates a protein ratio of the measured level in the red blood cell-enriched sample relative to the measured level in the separated plasma, and the produced protein profile comprises the calculated ratio having a value of at least 2:1.
The protein profile is based on a defined set of proteins, where the one or more proteins selected include basic fibroblast growth factor (FGF), CTACK, granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), hepatocyte growth factor (HGF), interferon alpha subtype α2 (IFN-α2), interferon gamma (IFN-γ), interleukin 12 p35 and p40 heterodimer (IL-12p70), IL-13, interleukin 12 p40 subunit (IL-12p40), IL-15, IL-16, IL-17A, IL-18, IL-1α, IL-1β, IL-2, interleukin 2 receptor alpha chain (IL-2ra), IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IP-10, LIF, M-CSF, MIG, MIP-1β, platelet-derived growth factor B chain homodimer (PDGF-BB), stromal cell-derived factor 1 (SDF-1α), tumor necrosis factor alpha (TNF-α), TNF-β, TNF-related apoptosis-inducing ligand (TRAIL), and vascular endothelial growth factor (VEGF).
Claims Coverage
Two independent claims are identified: clm-00001 and clm-00014. Each independent claim covers a method for producing a protein profile from leukodepleted red blood cell-enriched material, with the second independent claim further requiring a ratio relative to separated plasma with a minimum value.
Red blood cell-enriched secreted or released protein detection profile
A method of producing a protein profile by obtaining a blood sample, separating plasma from the blood sample to produce a cell pellet, leukodepleting the cell pellet to produce a red blood cell-enriched sample, incubating a small volume of the red blood cell-enriched sample in a medium, and detecting one or more proteins secreted or released from the red blood cell-enriched sample into the incubated medium, wherein the detected one or more proteins are selected from a specified group and wherein the protein profile comprises the detected one or more proteins in the incubated medium.
Red blood cell versus plasma protein ratio protein profile
A method of producing a protein profile by obtaining a blood sample, separating plasma from the blood sample to produce a cell pellet, leukodepletion of the cell pellet to produce a red blood cell-enriched sample, measuring the level of one or more proteins in the red blood cell-enriched sample and the level in the separated plasma, and calculating a protein ratio of the measured level in the red blood cell-enriched sample relative to the measured level in the separated plasma, wherein the produced protein profile comprises the calculated ratio having a value of at least 2:1, and wherein the one or more proteins are selected from a specified group.
Claim coverage centers on producing a protein profile from leukodepleted, red blood cell-enriched samples by detecting secreted or released proteins into a medium or by calculating a red blood cell-to-plasma protein ratio with a minimum threshold of at least 2:1, using a defined list of selectable proteins.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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