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Publication Number
US-11913964-B2
Publication Date
2024-02-27
Expiration Date
Abstract
Bead-based analytical assays suitable for detecting changes in the abundance of target analytes in biological samples are disclosed. In an embodiment, an assay involves incubating a sample with one or several beads that are capable of binding several distinct analytes in an amount sufficient for detection by mass spectrometry from a single bead.
Core Innovation
The invention relates to a method for analyzing a biological sample containing human cerebrospinal fluid (CSF). The method obtains a plurality of endogenous amyloid beta (A2) peptides by subjecting the biological sample to immunoprecipitation (IP), and the obtained endogenous A2 peptides are then analyzed using mass spectrometry by obtaining a mass spectrometry signal.
The method constrains the mass spectrometry signal acquisition to at least one endogenous CSF A2 peptide selected from A1-6, A1-7, A1-8, A1-9, A1-10, A1-21, A1-22, A1-23, A1-24, A1-25, A1-26, A1-31, and A1-32. Additional refinement specifies selected peptide subsets for which mass spectrometry signals are obtained, including A1-6 to A1-11 and A1-21 to A1-26.
Further refinement requires that biological samples be analyzed using mass spectrometry signals obtained from at least two endogenous CSF peptides that differ in a single terminal amino acid, with the peptides selected from the defined A2 sets. Additional refinement also specifies that the IP is performed using an antibody that specifically recognizes an endogenous A2 peptide, and that multiple antibody-based IP steps are used to obtain a first plurality and a second plurality of endogenous A2 peptides prior to mass spectrometry analysis.
Claims Coverage
The document provides three independent method claims. Across these independent claims, the core inventive coverage is concentrated on obtaining endogenous A2 peptides from human CSF by immunoprecipitation and analyzing selected endogenous A2 peptides by mass spectrometry signal acquisition from a defined peptide set.
Immunoprecipitation of endogenous A2 peptides from human CSF followed by mass spectrometry signal from selected endogenous CSF A2 peptides
Obtaining a plurality of endogenous amyloid beta (A2) peptides by subjecting a biological sample containing human cerebrospinal fluid (CSF) to immunoprecipitation (IP), and analyzing at least one of the plurality of endogenous A2 peptides using mass spectrometry, wherein the step of analyzing comprises obtaining a mass spectrometry signal from at least one endogenous CSF A2 peptide selected from A1-6, A1-7, A1-8, A1-9, A1-10, A1-21, A1-22, A1-23, A1-24, A1-25, A1-26, A1-31, and A1-32.
Antibody-specific immunoprecipitation of endogenous A2 followed by mass spectrometry signals for selected A2 peptides
Obtaining a plurality of endogenous amyloid beta (A2) peptides by subjecting a biological sample containing human cerebrospinal fluid (CSF) to immunoprecipitation (IP), and analyzing at least one of the plurality of endogenous A2 peptides using mass spectrometry, wherein analyzing comprises obtaining a mass spectrometry signal from at least one endogenous CSF A2 peptide selected from A1-6, A1-7, A1-8, A1-9, A1-10, A1-11, A1-21, A1-22, A1-23, A1-24, A1-25, A1-26, A1-31, and A1-32, and wherein the biological sample was previously subjected to IP using an antibody that specifically recognizes an endogenous A2 peptide.
First and second antibody immunoprecipitations to obtain separate endogenous A2 pluralities followed by mass spectrometry of selected A2 peptides
Obtaining a first plurality of endogenous amyloid beta (A2) peptides by subjecting a biological sample containing human cerebrospinal fluid (CSF) to immunoprecipitation (IP) using a first antibody, then obtaining a second plurality of endogenous A2 peptides by subjecting the biological sample to IP using a second antibody, and analyzing at least one of the first and the second pluralities using mass spectrometry, wherein the analyzing step comprises obtaining a mass spectrometry signal from at least one endogenous CSF A2 peptide selected from A1-6, A1-7, A1-8, A1-9, A1-10, A1-11, A1-21, A1-22, A1-23, A1-24, A1-25, A1-26, A1-31, and A1-32.
Across clm-00001, clm-00012, and clm-00015, the claimed methods share the combination of immunoprecipitation of endogenous A2 peptides from human CSF and mass spectrometry signal acquisition restricted to a defined set of endogenous CSF A2 peptides. Two independent claims further add limitations directed to antibody specificity for the IP step or to the use of first and second antibodies to generate separate A2 pluralities prior to analysis.
Stated Advantages
Not explicitly described in patent.
Documented Applications
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