# US 11,079,380 B2 · Optimizing diagnostics for galactofuranose containing antigens > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,079,380 B2 - Document ID: US11079380B2 - Status: Granted - Kind: B2 - Application number: 16/750,720 - Filed: 2020-01-23 - Published: 2020-07-16 - Granted: 2021-08-03 - Earliest claimed: 2009-11-23 - Assignees: The Johns Hopkins University - Applicants: The Johns Hopkins University - Inventors: Kieren Marr - Web page: https://patentgrep.com/patent/US11079380B2 ## CPC - G01N 33/56961: {Plant cells or fungi} - C07K 16/14: against material from fungi, algea or lichens - C07K 16/44: against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} - G01N 33/54366: {Apparatus specially adapted for solid-phase testing} - G01N 33/569: for microorganisms, e.g. protozoa, bacteria, viruses - G01N 33/56911: {Bacteria} - G01N 33/66: involving blood sugars, e.g. galactose - G01N 2400/02: involving antibodies to sugar part of glycoproteins - G01N 2469/10: Detection of antigens from microorganism in sample from host - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - 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) - 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) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract Disclosed herein are methods of detecting microbial infection in mammalian subjects comprising treatment of a sample and detection of galactofuranose (galF)-containing antigenic components utilizing monoclonal antibodies. The methods disclosed provide for pretreatment of biological samples, such as urine samples, to maximize detection of galF antigens and improvement of sensitivity of galF antigen detection assays. The methods include minimizing intelectin-1 binding to galF antigens and improvement of monoclonal antibody binding. The detection methods are useful for identifying the presence of microbial antigens related to bacterial, fungal, and parasitic pathogens, including Streptococcus pneumoniae, Aspergillus species, Fusarium species, Coccidioides species, Cryptococcus species, Histoplasma species, and Leishmania species. ## Claims ### Claim 1 A method for diagnosing a microbial infection in a biological sample from a mammalian subject suspected of having, having, or susceptible to having a microbial infection by detecting the presence of at least one polysaccharide comprising a galactofuranose residue in a biological sample of the mammalian subject, the method comprising: (a) treating the biological sample to inhibit human intelectin (hIntL) binding of galactofuranose residues present in the sample, wherein the treatment of the biological sample comprises contacting the biological sample with at least one ligand which binds directly to intelectin, or substrates which bind divalent cations with high affinity; (b) contacting the treated sample of (a) with at least one antibody specific for at least one polysaccharide comprising a galactofuranose residue in an effective amount to produce a detectable amount of antibody-polysaccharide complex, wherein the at least one antibody specific for at least polysaccharide comprising a galactofuranose residue is selected from the group consisting of monoclonal antibody 205 (MAb 205) comprising a variable heavy (VH) domain of SEQ ID NO:1 and a variable light (VL) domain of SEQ ID NO:2; monoclonal antibody 24 (MAb 24) comprising a VH domain of SEQ ID NO:3 and a VL domain of SEQ ID NO:4; monoclonal antibody 686 (MAb 686) comprising a VH domain of SEQ ID NO:5 and a VL domain of SEQ ID NO:6; monoclonal antibody 838 (MAb 838) comprising a VH domain of SEQ ID NO:7 and a VL domain of SEQ ID NO:8; and monoclonal antibody 476 (MAb 476) comprising a VH domain of SEQ ID NO:9 and a VL domain of SEQ ID NO:10; and (c) detecting the presence of at least one antibody-polysaccharide complex, wherein the detection of the presence of at least one antibody-polysaccharide complex is diagnostic of a microbial infection in a mammalian subject. ### Claim 2 (depends on 1) The method of claim 1, wherein in step (a) treating the sample comprises contacting the sample with a substrate. ### Claim 3 (depends on 2) The method of claim 2, wherein the substrate comprises a polyacrylamide resin. ### Claim 4 (depends on 2) The method of claim 2, wherein the substrate comprises a desalting column. ### Claim 5 (depends on 4) The method of claim 4, wherein the desalting column has a molecular weight cut off of less than 10 kDa. ### Claim 6 (depends on 4) The method of claim 4, wherein the desalting column has a molecular weight cut off of 7 kDa. ### Claim 7 A method for optimizing an assay for detecting the presence of at least one polysaccharide comprising a galF residue in a biological sample of the mammalian subject, the method comprising: (a) treating the biological sample comprising contacting the sample with a substrate which binds salts with high affinity, thereby inhibiting human intelectin (hIntL) binding of galF residues present in the sample; (b) contacting the treated sample of (a) with at least one antibody specific for at least one polysaccharide comprising a galF residue in an effective amount to produce a detectable amount of antibody-polysaccharide complex, wherein the at least one antibody specific for at least polysaccharide comprising a galactofuranose residue is selected from the group consisting of monoclonal antibody 205 (MAb 205) comprising a variable heavy (VH) domain of SEQ ID NO:1 and a variable light (VL) domain of SEQ ID NO:2; monoclonal antibody 24 (MAb 24) comprising a VH domain of SEQ ID NO:3 and a VL domain of SEQ ID NO:4; monoclonal antibody 686 (MAb 686) comprising a VH domain of SEQ ID NO:5 and a VL domain of SEQ ID NO:6; monoclonal antibody 838 (MAb 838) comprising a VH domain of SEQ ID NO:7 and a VL domain of SEQ ID NO:8; and monoclonal antibody 476 (MAb 476) comprising a VH domain of SEQ ID NO:9 and a VL domain of SEQ ID NO:10; and (c) detecting the presence of at least one antibody-polysaccharide complex. ### Claim 8 (depends on 7) The method of claim 7, wherein the substrate comprises a polyacrylamide resin. ### Claim 9 (depends on 7) The method of claim 7, wherein the substrate comprises a desalting column. ### Claim 10 (depends on 9) The method of claim 9, wherein the desalting column has a molecular weight cut off of less than 10 kDa. ### Claim 11 (depends on 9) The method of claim 9, wherein the desalting column has a molecular weight cut off of 7 kDa.