# US 2019/0376984 A1 · Methods for quantifying soluble amyloid beta and amyloid beta oligomers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0376984 A1 - Document ID: US20190376984A1 - Status: Published application - Kind: A1 - Application number: 16/436,244 - Filed: 2019-06-10 - Published: 2019-12-12 - Earliest claimed: 2018-06-08 - Inventors: Daniel Kidd; Janice Nickel - Web page: https://patentgrep.com/patent/US20190376984A1 ## CPC - G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease} - A61K 38/1716: {Amyloid plaque core protein} - C07K 16/18: against material from animals or humans - G01N 33/52: Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)} - C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues - G01N 2333/4709: Amyloid plaque core protein - G01N 2800/2821: Alzheimer - G01N 2800/50: Determining the risk of developing a disease - G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis - G01N 2800/56: Staging of a disease; Further complications associated with the disease - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Detection, diagnosis, and monitoring of Alzheimer's disease can be achieved with antibody-based detection methods directed to detect at least two regions of the amyloid beta peptide. The invention recognizes and relies on the observation that the steric availability of the carboxyl terminal epitope and the amino terminal epitope in an amyloid beta peptide differs based on whether the amyloid beta is monomeric or oligomeric. Due to oligomerization, the measured concentration of carboxyl termini that are available for binding represents the amount of monomeric amyloid beta in the sample; whereas the measured concentration of amino termini that are available for binding represents total amyloid beta (i.e., monomeric plus oligomeric) in the sample. Comparing the relative binding therefore can be used to characterize the population of soluble amyloid beta oligomers in the sample. The measurements are useful for early detection and subsequent monitoring of the development of Alzheimer's disease in a patient. ## Claims ### Claim 1 A method of characterizing amyloid beta, the method comprising: measuring a quantity of amyloid beta in a sample and measuring a quantity of a carboxyl-terminal portion of the amyloid beta available for binding in the sample; comparing the quantity of the carboxyl-terminal available for binding to the quantity of amyloid beta; and quantifying, based on the comparison, an amount of oligomeric amyloid beta in the sample. ### Claim 2 (depends on 1) The method of claim 1, wherein measuring the quantity of amyloid beta includes: contacting the sample with a reporter immunoreagent that binds to a mid-peptide region of the amyloid beta; contacting the sample with an immunoreagent that binds to an amino-terminal region of an amyloid beta peptide; and quantifying a detectable signal from a biomolecular interaction between amyloid beta peptides and the immunoreagents; measuring the quantity of the carboxyl-terminal portion of amyloid beta available for binding includes: contacting the sample with a second immunoreagent that binds to the carboxyl terminal; and measuring comprises quantifying a detectable signal from a biomolecular interaction between amyloid beta peptides and the immunoreagents. ### Claim 3 (depends on 1) The method of claim 1, wherein comparing comprises using an algorithm with the measured quantities as inputs, to generate an output comprising a quantified risk of amyloid beta mediated neurodegeneration; wherein the algorithm comprises: calculating a difference between the quantity of amyloid beta and the quantity of the carboxyl-terminal available for binding; calculating a ratio between the quantity of amyloid beta and the quantity of the carboxyl-terminal available for binding; or calculating a ratio between at least one of (i) the difference or sum between the quantity of amyloid beta and the quantity of the carboxyl-terminal available for binding and (ii) the quantity of amyloid beta. ### Claim 4 (depends on 2) The method of claim 2, wherein the reporter immunoreagent comprises 266, 6E10 or 4G8. ### Claim 5 (depends on 2) The method of claim 2, wherein the reporter immunoreagent is labeled with at least one of: an enzyme; a fluorophore, a chromophore; a chemiluminescent reagent; an affinity tag; or a radioisotopic reagent. ### Claim 6 (depends on 2) The method of claim 2, wherein the detectable signal is generated by an assay selected from the list consisting of colorimetric based Enzyme linked immunosorbent assay (ELISA) enzyme immunoassays (EIA), immunoblot assays, Western blot assays, immunoprecipitation, enzyme linked immunospot, antibody microarray assays, and/or immunohistochemistry assays (IHC), fluoroimmunoassays (FIA), fluorescent bead based immunoassay, flow cytometric assays, fluorescence-activated cell sorting (FACS), radioimmuno assays (RIA), immunocytochemical (ICH) assays, chemiluminescent assays, electrochemilumenescence (ECL) assays, chromatographic assays either by immunaffinity or post column signal generation. ### Claim 7 (depends on 1) The method of claim 1, further comprising: determining an amount of the monomeric amyloid beta based on the measured quantity of the carboxyl-terminal portion of the amyloid beta available for binding; determining an amount of the total (monomeric and oligomeric) amyloid beta based on the measured quantity of the amino-terminal portion of the amyloid beta available for binding; and providing a report that includes an amount of oligomeric amyloid beta in the sample, wherein the amount of oligomeric amyloid beta is the measured quantity of amyloid beta less the determined amount of monomeric amyloid beta. ### Claim 8 (depends on 7) The method of claim 7, wherein quantifying comprises determining a ratio between the amount of monomeric amyloid beta and total amyloid beta in the sample. ### Claim 9 (depends on 1) The method of claim 1, wherein the sample comprises at least one of blood, plasma, serum, or cerebrospinal fluid. ### Claim 10 (depends on 2) The method of claim 2, wherein the immunoreagent specifically binds to Aβ1, Aβ2, Aβ3, Aβ4, Aβ5, Aβ6, Aβ7, Aβ8, Aβ9, Aβ10, or Aβ11. ### Claim 11 (depends on 2) The method of claim 2, wherein the immunoreagent is 3D6, 1E8, MABN639 or AHP1252. ### Claim 12 (depends on 2) The method of claim 2, wherein the second immunoreagent specifically binds to Aβ37, Aβ38, Aβ39, Aβ40, Aβ41, or Aβ42. ### Claim 13 (depends on 2) The method of claim 2, wherein the second immunoreagent is anti-amyloid beta x-40 (2G3, MABN11) or anti-amyloid beta x-42 (MABN12, MABN13 or 05-831-1). ### Claim 14 (depends on 1) The method of claim 1, wherein one or both of the immunoreagents is a reporter antibody or a capture antibody. ### Claim 15 (depends on 1) The method of claim 1, further comprising assessing a subject's susceptibility to developing Alzheimer's disease; determining a stage of Alzheimer's disease; or determining a response to treatment of Alzheimer's disease or determining a treatment regimen. ### Claim 16 (depends on 2) The method of claim 2, further comprising treating the subject with an anti-idiotype antibody prior to contacting with the first and second immunoreagents, the anti-idiotype antibody comprising anti-solanuzumab or anti-bapineuzumab. ### Claim 17 A kit for characterizing amyloid beta in a sample, the kit comprising: an antibody specific for a carboxyl terminal epitope of amyloid beta, an antibody specific for an amino terminal epitope of amyloid beta, and an antibody specific for a central epitope of amyloid beta. ### Claim 18 (depends on 17) The kit of claim 17, wherein the carboxyl terminal antibody is end-specific for at least one of Aβ37, Aβ38, Aβ39, Aβ40, Aβ41, or Aβ42; and wherein the amino terminal antibody is end-specific for at least one of Aβ1, Aβ2, Aβ3, Aβ4, Aβ5, Aβ6, Aβ7, Aβ8, Aβ9 or Aβ10. ### Claim 19 A method of characterizing amyloid beta, the method comprising: measuring a quantity of amyloid beta in a sample and measuring a quantity of a carboxyl-terminal portion of the amyloid beta available for binding in the sample; assessing whether the quantity of the carboxyl-terminal available for binding meets a dilutional linearity acceptance criterion; and determining, based on the assessment, a relative amount of monomeric amyloid beta versus oligomeric amyloid beta in the sample. ### Claim 20 (depends on 19) The method of claim 19, wherein the carboxyl-terminal quantity meeting the dilutional linearity acceptance criterion indicates the sample contains mostly monomeric amyloid beta; and wherein the carboxyl-terminal quantity failing to meet the dilutional linearity acceptance criterion indicates the sample contains oligomeric amyloid beta.