# US 8,771,974 B2 · Methods of determining the presence and/or amount of a biomarker to determine the presence, identity, and/or severity of a lysosomal storage or neurological disorder > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,771,974 B2 - Document ID: US8771974B2 - Status: Granted - Kind: B2 - Application number: 13/550,106 - Filed: 2012-07-16 - Published: 2013-06-13 - Granted: 2014-07-08 - Earliest claimed: 2009-01-02 - Assignees: Biomarin Pharmaceutical Inc. - Applicants: Brett E. Crawford; Jillian R. Brown; Charles A. Glass; Jim R. Beitel; Robin M. Jackman - Inventors: Brett E. Crawford; Jillian R. Brown; Charles A. Glass; Jim R. Beitel; Robin M. Jackman - Web page: https://patentgrep.com/patent/US8771974B2 ## CPC - C12Q 1/527: involving lyase - C12Q 1/34: involving hydrolase - C12Q 1/40: involving amylase - G01N 33/6893: {related to diseases not provided for elsewhere} - G01N 2333/924: acting on glycosyl compounds (3.2) - G01N 2400/00: Assays, e.g. immunoassays or enzyme assays, involving carbohydrates - G01N 2800/042: Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism - G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - 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) ## Abstract Provided herein are methods of diagnosing or monitoring the treatment of abnormal glycan accumulation or a disorder associated with abnormal glycan accumulation. ## Claims ### Claim 1 A method of determining the presence of and/or measuring the amount of a biomarker to determine the presence, identity, and/or severity of the disease or condition in an individual having a disease or condition associated with abnormal glycan biosynthesis, degradation, or accumulation, the method comprising: (a) generating the biomarker comprising of one or more non-reducing end glycan residual compound(s), wherein the biomarker is generated by treating a population of glycans, in or isolated from a biological sample from the individual, with at least one digesting glycan enzyme(s), wherein prior to enzyme treatment, the biomarker is not present in abundance in samples from individuals with the disease or condition relative to individuals without the disease or condition, and (b) using an analytical instrument to detect the presence of and/or measure the amount of the biomarker produced and displaying or recording the presence of or a measure of a population of the biomarker; and (c) determining the presence, identity, and/or severity of the disease or condition utilizing the presence of and/or measure the amount of the biomarker; wherein the disease or condition is a lysosomal storage disorder or a neurological disorder; and wherein when the lysosomal storage disorder is an MPS disorder, then the digesting glycan enzyme is not a lyase. ### Claim 2 (depends on 1) The method of claim 1, wherein the disease or disorder is caused by an abnormally functioning glycan degradation enzyme and wherein the abnormally functioning glycan degradation enzyme and the normally functioning glycan degradation enzyme are of the same type. ### Claim 3 (depends on 1) The method of claim 1, wherein the abnormal glycan accumulation comprises the accumulation of abnormal amounts of normal glycans. ### Claim 4 (depends on 1) The method of claim 1, wherein the abnormal glycan accumulation comprises the accumulation of abnormal amounts of abnormal glycans. ### Claim 5 (depends on 1) The method of claim 1, wherein the normally functioning glycan degradation enzyme is a glycosidase, sulfatase, phosphorylase, deacetylase, or a combination thereof. ### Claim 6 (depends on 5) The method of claim 5, wherein the normally functioning glycan degradation enzyme is a glycosidase selected from an exo-glycosidase and an endo-glycosidase. ### Claim 7 (depends on 6) The method of claim 6 wherein the glycosidase is an exo-glycosidase selected from the group consisting of a galactosidase, and a glucuronidase. ### Claim 8 (depends on 6) The method of claim 6, wherein the glycan residual compound is a monosaccharide. ### Claim 9 (depends on 5) The method of claim 5, wherein the glycan residual compound is sulfate, phosphate, acetate, or a combination thereof. ### Claim 10 (depends on 1) The method of claim 1, further comprising purifying a biological sample prior to transforming a glycan thereof wherein the process of purifying a biological sample comprises removing monosaccharides therefrom, removing sulfates therefrom, removing phosphates therefrom, removing acetate therefrom, or a combination thereof. ### Claim 11 (depends on 1) The method of claim 1, wherein transforming a glycan of a biological sample with a normally functioning glycan degradation enzyme comprises transforming a glycan of a biological sample with a plurality of normally functioning glycan degradation enzymes concurrently, sequentially, or a combination thereof. ### Claim 12 (depends on 1) The method of claim 1, wherein the disorder associated with an abnormal glycan accumulation is Alzheimer's disease, Parkinson's disease, MPS I, MPS II, MPS IIIA, MPS IVA, MPS VI, Gaucher disease, Krabbe disease, Pompe disease, Sandhoff disease, Tay-Sachs disease, or Fabry Disease. ### Claim 13 (depends on 1) The method of claim 1, wherein determining whether the amount of liberated glycan residue is abnormal comprises labeling the glycan residue with a detectable label and measuring the amount of labeled glycan residue with an analytical instrument using UV-Vis spectroscopy, IR spectroscopy, mass spectrometry, or a combination thereof. ### Claim 14 (depends on 13) The method of claim 13, wherein the detectable label is a mass label, a radioisotope label, a fluorescent label, a chromophore label, or affinity label. ### Claim 15 (depends on 1) The method of claim 1, wherein prior to measuring the amount of a population of non-reducing end glycan residual compounds, the non-reducing end glycan residual compounds are labeled with a detectable label and wherein the amount of liberated glycan is measured using UV-Vis spectroscopy, IR spectroscopy, mass spectrometry, or a combination thereof. ### Claim 16 (depends on 15) The method of claim 15, wherein the detectable label is a mass label, a radioisotope label, a fluorescent label, a chromophore label, or affinity label. ### Claim 17 A method monitoring the treatment of a disorder associated with the abnormal degradation, biosynthesis and/or accumulation of glycans, the method comprising: (a) following administration of an agent for treating a disorder associated with the abnormal degradation, biosynthesis and/or accumulation of glycans to an individual in need thereof, using an analytical instrument to measure the amount of a population of a biomarker comprising non-reducing end glycan residual compounds present in a transformed biological sample, the biomarker being generated by treating a population of glycans, in or isolated from a biological sample from the individual, with at least one digesting glycan enzyme(s), wherein prior to enzyme treatment, the biomarker is not present in abundance in samples from individuals with the disease or condition relative to individuals without the disease or condition, and (b) determining whether or not the amount of biomarker has decreased or increased at a slower rate compared to the amount or rate of increase prior to administration of the agent for treating a disorder associated with the abnormal degradation, biosynthesis and/or accumulation of glycans; and wherein the disorder is a lysosomal disorder or a neurological disorder. ### Claim 18 (depends on 17) The method of claim 17, wherein the normally functioning glycan degradation enzyme is a glycosidase, sulfatase, phosphorylase, deacetylase, or a combination thereof. ### Claim 19 (depends on 18) The method of claim 18, wherein the normally functioning glycan degradation enzyme is a glycosidase selected from an exo-glycosidase and an endo-glycosidase. ### Claim 20 (depends on 17) The method of claim 17, wherein the glycan residual compound is a monosaccharide, sulfate, phosphate, acetate, or a combination thereof. ### Claim 21 (depends on 17) The method of claim 17, wherein transforming a glycan of a biological sample with a normally functioning glycan degradation enzyme comprises transforming a glycan of a biological sample with a plurality of normally functioning glycan degradation enzymes concurrently, sequentially, or a combination thereof.