Protein glycosylation in urine as a biomarker of diseases

  • Author Footnotes
    # These authors contribute equally to this work.
    Mingming Xu
    # These authors contribute equally to this work.
    Center for Clinical Mass Spectrometry, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, China
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  • Author Footnotes
    # These authors contribute equally to this work.
    Arthur Yang
    # These authors contribute equally to this work.
    Center for Clinical Mass Spectrometry, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, China
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  • Jun Xia
    Clinical Laboratory Center, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang, China
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  • Junhong Jiang
    Department of Pulmonary and Critical Care Medicine, Dushu Lake Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Chun-Feng Liu
    Department of Neurology and Clinical Research Center of Neurological Disease, The Second Affiliated Hospital of Soochow University, Suzhou, China
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  • Zhenyu Ye
    Department of General Surgery, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Junfeng Ma
    Reprint requests: Junfeng Ma: Room GD9, Pre-Clinical Science Building, 3900 Reservoir Road NW, Washington DC 20007
    Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Georgetown University, Washington, District of Columbia
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  • Shuang Yang
    Reprint requests: Shuang Yang: 2107 Yunxuan Bld, 199 Ren'Ai Lu, Suzhou Industrial Park, Suzhou, Jiangsu 215123, China
    Center for Clinical Mass Spectrometry, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, China
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  • Author Footnotes
    # These authors contribute equally to this work.
Published:August 08, 2022DOI:


      Human body fluids have become an indispensable resource for clinical research, diagnosis and prognosis. Urine is widely used to discover disease-specific glycoprotein biomarkers because of its recurrently non-invasive collection and disease-indicating properties. While urine is an unstable fluid in that its composition changes with ingested nutrients and further as it is excreted through micturition, urinary proteins are more stable and their abnormal glycosylation is associated with diseases. It is known that aberrant glycosylation can define tumor malignancy and indicate disease initiation and progression. However, a thorough and translational survey of urinary glycosylation in diseases has not been performed. In this article, we evaluate the clinical applications of urine, introduce methods for urine glycosylation analysis, and discuss urine glycoprotein biomarkers. We emphasize the importance of mining urinary glycoproteins and searching for disease-specific glycosylation in various diseases (including cancer, neurodegenerative diseases, diabetes, and viral infections). With advances in mass spectrometry-based glycomics/glycoproteomics/glycopeptidomics, characterization of disease-specific glycosylation will optimistically lead to the discovery of disease-related urinary biomarkers with better sensitivity and specificity in the near future.


      A1BG (alpha-1B-glycoprotein), A4GALT (lactosylceramide 4-alpha-galactosyltransferase), AAK1 (AP2-associated protein kinase 1), AD (Alzheimer's disease), ADAM (metallopeptidase), AF (amniotic fluid), AFP (alpha-fetoprotein), AGE (advanced glycation end product), AGP (alpha-1-acid glycoprotein), ALS (amyotrophic lateral sclerosis), AMYP (alpha-amylase), APOA1 (apolipoprotein A1), APOC3 (apolipoprotein C3), AUC (area under curve), BALF (bronchoalveolar lavage fluid), BBB (blood-brain barrier), BM (breast milk), CA125 (cancer antigen 125), CA19-9 (cancer antigen 19-9), CCDC132 (coiled-coil domain-containing protein 132), CE (capillary electrophoresis), CEA (carcinoembryonic antigen), CNS (central nervous system), CoA (coenzyme A), Cp (ceruloplasmin), CSF (cerebrospinal fluid), DAC (duodenal adenocarcinomas), DNA (deoxyribonucleic acid), DNP (diabetic nephropathy), EIA (enzyme immunoassay), ELISA (enzyme-linked immunoassay), ER (endoplasmic reticulum), EV (extracellular vesicle), FTD (frontotemporal dementia), FUS/TLS (fused in sarcoma/translated in liposarcoma), FUT8 (fucosyltransferase 8), GALNT (polypeptide N-acetylgalactosaminyltransferase), GP2-1 (glycoprotein gp2-1), HA (hemagglutinin), HBF (human body fluid), HBFP (human body fluid proteome), HBP (hexosamine biosynthesis pathway), HCC (hepatocellular carcinomas), HCV (hepatitis C virus), HD (Huntington's disease), HDAC6 (histone deacetylase 6), HILIC (hydrophilic interaction liquid chromatography), HPA (human protein atlas), IAV (influenza A virus), IBV (influenza B virus), IGF-2 (insulin-like growth factor 2), IgG (immunoglobulin), IGHV3-15 (immunoglobulin heavy variable 3-15), IGHV3-30 (immunoglobulin heavy variable 3-30), IGHV4-28 (immunoglobulin heavy variable 4-28), IGKJ1 (immunoglobulin kappa joining 1), IGKV1-8 (immunoglobulin kappa variable 1-8), IGKV3D-15 (immunoglobulin kappa variable 3D-15), IGLJ3 (immunoglobulin lambda joining 3), IGLV4-69 (immunoglobulin lambda variable 4-69), IL6 (interleukin-6), ITIH4 (inter-alpha-trypsin inhibitor heavy chain H4), LC (liquid chromatography), LPS (lipopolysaccharide), LRRK2 (leucine-rich repeat serine/threonine-protein kinase 2), LYVE1 (lymphatic vessel endothelial hyaluronic acid receptor 1), MALDI (matrix-assisted laser desorption/ionization), MCI (mild cognitive impairment), MMP14 (matrix metalloproteinase 14), MND (motor neuron disease), MS (mass spectrometry), MWCO (molecular-weight-cutoff filter), NA (neuraminidase), NAF (nipple aspirate fluid), ND (neurodegenerative disease), NMP22 (nuclear matrix protein 22), NR4A2 (nuclear receptor-related factor 1), NSCLC (non-small cell lung cancer), NSE (neuron specific enolase), OGA (O-GlcNAcase), O-GIG (O-Glycopeptide immobilization for O-glycosylated peptide enrichment), OGT (O-GlcNAc transferase SQSTM1, sequestosome-1 p75NTR, p75 neurotrophin receptor), PAAC (prostate acinar adenocarcinomas), PC (pancreatic cancer), PCR (polymerase chain reaction), PD (Parkinson's disease), PDAC (pancreatic ductal adenocarcinoma), PNGase F (peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase), PRSS1 (serine protease 1), PSA (prostate-specific antigen), PTM (post-translational modification), REG1A (lithostathine-1-alpha), SCC (nipped-B-like protein), SCLC (small cell lung cancer), SERPINA1 (alpha-1-antitrypsin), SHROOM1 (protein shroom1), SLC14A1 (urea transporter 1), sLeX (sialyl lewis X), SqCC (squamous cell carcinoma), T1DM (type 1 diabetes mellitus), T2DM (type 2 diabetes mellitus), TACR3 (neuromedin-K receptor), TCC (transitional cell cancer), TDP-43 (TAR DNA-binding protein 43), TF (tear fluid), TFF1 (trefoil factor 1), TNF (tumor necrosis factor-alpha), Trf (serotransferrin), TSG (tumor-specific glycosylation), TTR (transthyretin), UC (ultracentrifugation), UDP-GlcNAc (uridine diphosphate-N-acetylglucosamine), UPK3B (uroplakin-3b)
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        • Hu S
        • Loo JA
        • Wong DT.
        Human body fluid proteome analysis.
        Proteomics. 2006; 6: 6326-6353
        • Zhang Z
        • Bast Jr., RC
        • Yu Y
        Three biomarkers identified from serum proteomic analysis for the detection of early stage ovarian cancer.
        Cancer Res. 2004; 64: 5882-5890
        • Zinkin NT
        • Grall F
        • Bhaskar K
        Serum proteomics and biomarkers in hepatocellular carcinoma and chronic liver disease.
        Clin Cancer Res. 2008; 14: 470-477
        • Su D
        • Li W
        • Xu Q
        New metabolites of acteoside identified by ultra-performance liquid chromatography/quadrupole-time-of-flight MSE in rat plasma, urine, and feces.
        Fitoterapia. 2016; 112: 45-55
        • Ginsberg JM
        • Chang BS
        • Matarese RA
        • Garella S.
        Use of single voided urine samples to estimate quantitative proteinuria.
        N Engl J Med. 1983; 309: 1543-1546
        • Ritz E
        • Orth SR.
        Nephropathy in patients with type 2 diabetes mellitus.
        N Engl J Med. 1999; 341: 1127-1133
        • Currie G
        • Delles C.
        Proteinuria and its relation to cardiovascular disease.
        Int J Nephrol Renovasc Dis. 2014; 7: 13-24
        • Soloway MS
        • Briggman JV
        • Carpinito GA
        Use of a new tumor marker, urinary NMP22, in the detection of occult or rapidly recurring transitional cell carcinoma of the urinary tract following surgical treatment.
        J Urol. 1996; 156: 363-367
        • Ma J
        • Li Y
        • Hou C
        • Wu C.
        O-GlcNAcAtlas: a database of experimentally identified O-GlcNAc sites and proteins.
        Glycobiology. 2021; 31: 719-723
        • Mian C
        • Lodde M
        • Haitel A
        Comparison of the monoclonal UBC-ELISA test and the NMP22 ELISA test for the detection of urothelial cell carcinoma of the bladder.
        Urology. 2000; 55: 223-226
        • Moonen P
        • Kiemeney L
        • Witjes J.
        Urinary NMP22® BladderChek® test in the diagnosis of superficial bladder cancer.
        Eur Urol. 2005; 48: 951-956
        • Dotz V
        • Visconti A
        • Lomax-Browne HJ
        O-and N-glycosylation of serum immunoglobulin A is associated with IgA nephropathy and glomerular function.
        J Am Soc Nephrol. 2021; 32: 2455-2465
        • Kanamori-Katayama M
        • Kaiho A
        • Ishizu Y
        LRRN4 and UPK3B are markers of primary mesothelial cells.
        PLoS One. 2011; 6: 1-8
        • Rudat C
        • Grieskamp T
        • Röhr C
        Upk3b is dispensable for development and integrity of urothelium and mesothelium.
        PLoS One. 2014; 9: 1-10
        • Sidoux-Walter F
        • Lucien N
        • Nissinen R
        Molecular heterogeneity of the Jk(null) phenotype: expression analysis of the Jk(S291P) mutation found in Finns.
        Blood. 2000; 96: 1566-1573
        • Shigemoto R
        • Yokota Y
        • Tsuchida K
        • Nakanishi S.
        Cloning and expression of a rat neuromedin K receptor cDNA.
        J Biol Chem. 1990; 265: 623-628
        • Topaloglu AK
        • Reimann F
        • Guclu M
        TAC3 and TACR3 mutations in familial hypogonadotropic hypogonadism reveal a key role for Neurokinin B in the central control of reproduction.
        Nat Genet. 2009; 41: 354-358
        • MAdS Silva
        • Lenz B
        • Rotter A
        Neurokinin3 receptor as a target to predict and improve learning and memory in the aged organism.
        Proc Natl Acad Sci USA. 2013; 110: 15097-15102
        • Snyder H
        • Wolozin B.
        Pathological proteins in Parkinson's disease.
        J Mol Neurosci. 2004; 24: 425-442
        • Handley RR
        • Reid SJ
        • Brauning R
        Brain urea increase is an early Huntington's disease pathogenic event observed in a prodromal transgenic sheep model and HD cases.
        Proc Natl Acad Sci USA. 2017; 114: E11293-E1E302
        • Jones AC
        • Pinki F
        • Stewart GS
        • Costello DA.
        Inhibition of urea transporter (UT)-B modulates LPS-induced inflammatory responses in BV2 microglia and N2a neuroblastoma cells.
        Neurochem Res. 2021; 46: 1322-1329
        • Schjoldager KT
        • Narimatsu Y
        • Joshi HJ
        • Clausen H.
        Global view of human protein glycosylation pathways and functions.
        Nat Rev Mol Cell Biol. 2020; 21: 729-749
        • Varki A.
        Biological roles of glycans.
        Glycobiology. 2017; 27: 3-49
        • Hart GW
        • Housley MP
        • Slawson C.
        Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins.
        Nature. 2007; 446: 1017-1022
        • Hart GW.
        Nutrient regulation of signaling and transcription.
        J Biol Chem. 2019; 294: 2211-2231
        • Chatham JC
        • Zhang J
        • Wende AR.
        Role of O-linked N-acetylglucosamine protein modification in cellular (patho) physiology.
        Physiol Rev. 2021; 101: 427-493
        • Halim A
        • Nilsson J
        • Rüetschi U
        • Hesse C
        • Larson G.
        Human urinary glycoproteomics; attachment site specific analysis of N-and O-linked glycosylations by CID and ECD.
        Mol Cell Proteomics. 2012; 11: 1-17
        • Sun S
        • Hu Y
        • Ao M
        N-GlycositeAtlas: a database resource for mass spectrometrybased human N-linked glycoprotein and glycosylation site mapping.
        Clin Proteomics. 2019; 16: 1-11
        • Thomas CE
        • Sexton W
        • Benson K
        • Sutphen R
        • Koomen J.
        Urine collection and processing for protein biomarker discovery and quantification.
        Cancer Epidemiol Biomarkers Prev. 2010; 19: 953-959
        • Gonzales PA
        • Zhou H
        • Pisitkun T
        Isolation and purification of exosomes in urine. The urinary proteome.
        Springer, 2010: 89-99
        • Street J
        • Koritzinsky E
        • Glispie D
        • Star R
        • Yuen P.
        Urine exosomes: an emerging trove of biomarkers.
        Adv Clin Chem. 2017; 78: 103-122
        • Pisitkun T
        • Shen R-F
        • Knepper MA.
        Identification and proteomic profiling of exosomes in human urine.
        Proc Natl Acad Sci USA. 2004; 101: 13368-13373
        • Zou G
        • Benktander JD
        • Gizaw ST
        • Gaunitz S
        • Novotny MV.
        Comprehensive analytical approach toward glycomic characterization and profiling in urinary exosomes.
        Anal Chem. 2017; 89: 5364-5372
        • Song W
        • Zhou X
        • Benktander JD
        In-depth compositional and structural characterization of N-glycans derived from human urinary exosomes.
        Anal Chem. 2019; 91: 13528-13537
        • Brown CJ
        • Gaunitz S
        • Wang Z
        Glycoproteomic analysis of human urinary exosomes.
        Anal Chem. 2020; 92: 14357-14365
        • Van Deun J
        • Mestdagh P
        • Sormunen R
        The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling.
        J Extracell Vesicles. 2014; 3: 1-14
        • Nakai W
        • Yoshida T
        • Diez D
        A novel affinity-based method for the isolation of highly purified extracellular vesicles.
        Sci Rep. 2016; 6: 1-11
        • Merchant ML
        • Rood IM
        • Deegens JK
        • Klein JB.
        Isolation and characterization of urinary extracellular vesicles: implications for biomarker discovery.
        Nat Rev Nephrol. 2017; 13: 731-749
        • Yoshikawa H
        • Hirano A
        • Arakawa T
        • Shiraki K.
        Effects of alcohol on the solubility and structure of native and disulfide-modified bovine serum albumin.
        Int J Biol Macromol. 2012; 50: 1286-1291
        • Kammeijer GSM
        • Nouta J
        • de la Rosette JJMCH
        • de Reijke TM
        • Wuhrer M.
        An in-depth glycosylation assay for urinary prostate-specific antigen.
        Anal Chem. 2018; 90: 4414-4421
        • Li H
        • Patel V
        • DiMartino SE
        • Froehlich JW
        • Lee RS.
        An in-depth comparison of the pediatric and adult urinary N-glycomes.
        Mol Cell Proteomics. 2020; 19: 1767-1776
        • Li H
        • Kostel SA
        • DiMartino SE
        Uromodulin isolation and its N-glycosylation analysis by nanoLC-MS/MS.
        J Proteome Res. 2021; 20: 2662-2672
        • Kreunin P
        • Zhao J
        • Rosser C
        Bladder cancer associated glycoprotein signatures revealed by urinary proteomic profiling.
        J Proteome Res. 2007; 6: 2631-2639
        • Yang N
        • Feng S
        • Shedden K
        Urinary glycoprotein biomarker discovery for bladder cancer detection using LC/MS-MS and label-free quantification.
        Clin Cancer Res. 2011; 17: 3349-3359
        • Martin DT
        • Shen H
        • Steinbach-Rankins JM
        Glycoprotein-130 expression is associated with aggressive bladder cancer and is a potential therapeutic target.
        Mol Cancer Ther. 2019; 18: 413-420
        • Yang S
        • Clark D
        • Liu Y
        • Li S
        • Zhang H
        High-throughput analysis of N-glycans using AutoTip via glycoprotein immobilization.
        Sci Rep. 2017; 7: 1-11
        • Yang S
        • Hu Y
        • Sokoll L
        • Zhang H.
        Simultaneous quantification of N-and O-glycans using a solid-phase method.
        Nat Protoc. 2017; 12: 1229-1244
        • Chen S-Y
        • Dong M
        • Yang G
        Glycans, glycosite, and intact glycopeptide analysis of Nlinked glycoproteins using liquid handling systems.
        Anal Chem. 2020; 92: 1680-1686
        • Packer NH
        • Lawson MA
        • Jardine DR
        • Redmond JW.
        A general approach to desalting oligosaccharides released from glycoproteins.
        Glycoconj J. 1998; 15: 737-747
        • Jensen PH
        • Karlsson NG
        • Kolarich D
        • Packer NH.
        Structural analysis of N-and O-glycans released from glycoproteins.
        Nat Protoc. 2012; 7: 1299-1310
        • Yang S
        • Li Y
        • Shah P
        • Zhang H.
        Glycomic analysis using glycoprotein immobilization for glycan extraction.
        Anal Chem. 2013; 85: 5555-5561
        • Sun X
        • Tao L
        • Yi L
        N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database.
        J Pharm Anal. 2017; 7: 87-94
        • Yang S
        • Yuan W
        • Yang W
        Glycan analysis by isobaric aldehyde reactive tags and mass spectrometry.
        Anal Chem. 2013; 85: 8188-8195
        • Yang S
        • Wang M
        • Chen L
        QUANTITY: an isobaric tag for quantitative glycomics.
        Sci Rep. 2015; 5: 1-10
        • Alvarez-Manilla G
        • Warren NL
        • Abney T
        Tools for glycomics: relative quantitation of glycans by isotopic permethylation using 13CH3I.
        Glycobiology. 2007; 17: 677-687
        • Ruhaak L
        • Zauner G
        • Huhn C
        Glycan labeling strategies and their use in identification and quantification.
        Anal Bioanal Chem. 2010; 397: 3457-3481
        • Hao P
        • Ren Y
        • Alpert AJ
        • Sze SK.
        Detection, evaluation and minimization of nonenzymatic deamidation in proteomic sample preparation.
        Mol Cell Proteomics. 2011; 10: 1-11
        • Cao L
        • Diedrich JK
        • Ma Y
        Global site-specific analysis of glycoprotein N-glycan processing.
        Nat Protoc. 2018; 13: 1196-1212
        • Zhang H
        • Li X-j
        • Martin DB
        • Aebersold R.
        Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry.
        Nat Biotechnol. 2003; 21: 660-666
        • Kaji H
        • Saito H
        • Yamauchi Y
        Lectin affinity capture, isotope-coded tagging and mass spectrometry to identify N-linked glycoproteins.
        Nat Biotechnol. 2003; 21: 667-672
        • Zauner G
        • Deelder AM
        • Wuhrer M.
        Recent advances in hydrophilic interaction liquid chromatography (HILIC) for structural glycomics.
        Electrophoresis. 2011; 32: 3456-3466
        • Li J
        • Zhang J
        • Xu M
        Advances in glycopeptide enrichment methods for the analysis of protein glycosylation over the past decade.
        J Sep Sci. 2022; 45: 3169-3186
        • Riley NM
        • Bertozzi CR
        • Pitteri SJ.
        A pragmatic guide to enrichment strategies for mass spectrometry-based glycoproteomics.
        Mol Cell Proteomics. 2021; 20: 1-29
        • Ma J
        • Hou C
        • Wu C.
        Demystifying the O-GlcNAc code: a systems view.
        Chem Rev. 2022; 1: 1-43
        • Vainauskas S
        • Guntz H
        • McLeod E
        A broad-specificity O-glycoprotease that enables improved analysis of glycoproteins and glycopeptides containing intact complex O-glycans.
        Anal Chem. 2022; 94: 1060-1069
        • Yang S
        • Onigman P
        • Wu WW
        Deciphering protein O-glycosylation: solid-phase chemoenzymatic cleavage and enrichment.
        Anal Chem. 2018; 90: 8261-8269
        • Yang S
        • Wu WW
        • Shen R
        Optimization of O-GIG for O-glycopeptide characterization with sialic acid linkage determination.
        Anal Chem. 2020; 92: 10946-10951
        • Malaker SA
        • Pedram K
        • Ferracane MJ
        The mucin-selective protease StcE enables molecular and functional analysis of human cancer-associated mucins.
        Proc Natl Acad Sci USA. 2019; 116: 7278-7287
        • Yang S
        • Wang PG
        Method development of glycoprotein biomarkers for cancers.
        Bioanalysis. 2017; 9: 903-906
        • Xu M-m
        • Zhou M-t
        • Li S-w
        • Zhen X-c
        • Yang S
        Glycoproteins as diagnostic and prognostic biomarkers for neurodegenerative diseases: a glycoproteomic approach.
        J Neurosci Res. 2021; 99: 1308-1324
        • Thompson A
        • Schäfer J
        • Kuhn K
        Tandem mass tags:  a novel quantification strategy for comparative analysis of complex protein mixtures by MS/MS.
        Anal Chem. 2003; 75: 1895-1904
        • Gan CS
        • Chong PK
        • Pham TK
        • Wright PC.
        Technical, experimental, and biological variations in isobaric tags for relative and absolute quantitation (iTRAQ).
        J Proteome Res. 2007; 6: 821-827
        • Ren Y
        • He Y
        • Lin Z
        Reagents for isobaric labeling peptides in quantitative proteomics.
        Anal Chem. 2018; 90: 12366-12371
        • Li J
        • Van Vranken JG
        • Pontano Vaites L
        TMTpro reagents: a set of isobaric labeling mass tags enables simultaneous proteome-wide measurements across 16 samples.
        Nat Methods. 2020; 17: 399-404
        • Alley Jr, WR
        • Mechref Y
        • Novotny MV
        Characterization of glycopeptides by combining collision-induced dissociation and electron-transfer dissociation mass spectrometry data.
        Rapid Commun Mass Spectrom. 2009; 23: 161-170
        • Riley NM
        • Malaker SA
        • Driessen MD
        • Bertozzi CR.
        Optimal dissociation methods differ for N- and O-glycopeptides.
        J Proteome Res. 2020; 19: 3286-3301
        • Yang S
        • Wang Y
        • Mann M
        Improved online LC-MS/MS identification of O-glycosites by EThcD fragmentation, chemoenzymatic reaction, and SPE enrichment.
        Glycoconj J. 2021; 38: 145-156
        • Bern M
        • Kil YJ
        • Becker C.
        Byonic: advanced peptide and protein identification software.
        Curr Protoc Bioinformatics. 2012; (Chapter 13:Unit13.20-Unit13.20)
        • Maxwell E
        • Tan Y
        • Tan Y
        GlycReSoft: a software package for automated recognition of glycans from LC/MS data.
        PLoS One. 2012; 7: 1-11
        • Ramezani A
        • Devaney JM
        • Cohen S
        Circulating and urinary micro RNA profile in focal segmental glomerulosclerosis: a pilot study.
        Eur J Clin Invest. 2015; 45: 394-404
        • Bishnoi M
        • Chopra K
        • Kulkarni SK.
        Possible anti-oxidant and neuroprotective mechanisms of zolpidem in attenuating typical anti-psychotic-induced orofacial dyskinesia-a biochemical and neurochemical study.
        Prog Neuropsychopharmacol Biol Psychiatry. 2007; 31: 1130-1138
        • Nichkova M
        • Wynveen PM
        • Marc DT
        • Huisman H
        • Kellermann GH.
        Validation of an ELISA for urinary dopamine: applications in monitoring treatment of dopamine-related disorders.
        J Neurochem. 2013; 125: 724-735
        • Sjöstedt E
        • Zhong W
        • Fagerberg L
        An atlas of the protein-coding genes in the human, pig, and mouse brain.
        Science. 2020; 367: 1-16
        • Sjöstedt E
        • Fagerberg L
        • Hallström BM
        Defining the human brain proteome using transcriptomics and antibody-based profiling with a focus on the cerebral cortex.
        PLoS One. 2015; 10: 1-20
        • An M
        • Gao Y
        Urinary biomarkers of brain diseases.
        Genomics Proteomics Bioinformatic. 2015; 13: 345-354
        • Chandler K
        • Goldman R.
        Glycoprotein disease markers and single protein-omics.
        Mol Cell Proteomics. 2013; 12: 836-845
        • Stanley P
        • Taniguchi N
        • Aebi M
        • et al.
        in: Varki A Cummings RD Esko JD Essentials of glycobiology. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY2017
        • Poruk KE
        • Gay DZ
        • Brown K
        The clinical utility of CA 19-9 in pancreatic adenocarcinoma: diagnostic and prognostic updates.
        Curr Mol Med. 2013; 13: 340-351
        • Radon TP
        • Massat NJ
        • Jones R
        Identification of a three-biomarker panel in urine for early detection of pancreatic adenocarcinoma.
        Clin Cancer Res. 2015; 21: 3512-3521
        • Jackson DG
        • Prevo R
        • Clasper S
        • Banerji S.
        LYVE-1, the lymphatic system and tumor lymphangiogenesis.
        Trends Immunol. 2001; 22: 317-321
        • Nightingale TD
        • Frayne ME
        • Clasper S
        • Banerji S
        • Jackson DG.
        A mechanism of sialylation functionally silences the hyaluronan receptor LYVE-1 in lymphatic endothelium.
        J Biol Chem. 2009; 284: 3935-3945
        • Chen R
        • Jiang X
        • Sun D
        Glycoproteomics analysis of human liver tissue by combination of multiple enzyme digestion and hydrazide chemistry.
        J Proteome Res. 2009; 8: 651-661
        • Bollineni RC
        • Koehler CJ
        • Gislefoss RE
        • Anonsen JH
        • Thiede B.
        Large-scale intact glycopeptide identification by Mascot database search.
        Sci Rep. 2018; 8: 1-13
        • Yin X
        • Bern M
        • Xing Q
        Glycoproteomic analysis of the secretome of human endothelial cells.
        Mol Cell Proteomics. 2013; 12: 956-978
        • Banerji S
        • Ni J
        • Wang S-X
        LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.
        J Cell Biol. 1999; 144: 789-801
        • De Caro AM
        • Adrich Z
        • Fournet B
        N-terminal sequence extension in the glycosylated forms of human pancreatic stone protein. The 5-oxoproline N-terminal chain is O-glycosylated on the 5th amino acid residue.
        Biochim Biophys Acta. 1989; 994: 281-284
        • Fagerberg L
        • Hallström BM
        • Oksvold P
        Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics.
        Mol Cell Proteomics. 2014; 13: 397-406
        • Astrosini C
        • Roeefzaad C
        • Dai YY
        REG1A expression is a prognostic marker in colorectal cancer and associated with peritoneal carcinomatosis.
        Int J Cancer. 2008; 123: 409-413
        • Im S
        • Yoo C
        • Jung J-H
        Reduced expression of TFF1 and increased expression of TFF3 in gastric cancer: correlation with clinicopathological parameters and prognosis.
        Int J Med Sci. 2013; 10: 133-140
        • Wang W
        • Li Z
        • Wang J
        A functional polymorphism in TFF1 promoter is associated with the risk and prognosis of gastric cancer.
        Int J Cancer. 2018; 142: 1805-1816
        • Sankpal N
        • Moskaluk C
        • Hampton G
        • Powell S.
        Overexpression of CEBPβ correlates with decreased TFF1 in gastric cancer.
        Oncogene. 2006; 25: 643-649
        • Porterfield M
        • Zhao P
        • Han H
        Discrimination between adenocarcinoma and normal pancreatic ductal fluid by proteomic and glycomic analysis.
        J Proteome Res. 2014; 13: 395-407
        • Kammeijer GS
        • Nouta J
        • de la Rosette JJ
        • de Reijke TM
        • Wuhrer M.
        An in-depth glycosylation assay for urinary prostate-specific antigen.
        Anal Chem. 2018; 90: 4414-4421
        • Wang X
        • Chen J
        • Li QK
        Overexpression of α (1, 6) fucosyltransferase associated with aggressive prostate cancer.
        Glycobiology. 2014; 24: 935-944
        • Höti N
        • Yang S
        • Hu Y
        Overexpression of α (1, 6) fucosyltransferase in the development of castration-resistant prostate cancer cells.
        Prostate Cancer Prostatic Dis. 2018; 21: 137-146
        • Stura EA
        • Muller BH
        • Bossus M
        Crystal structure of human prostate-specific antigen in a sandwich antibody complex.
        J Mol Biol. 2011; 414: 530-544
        • Clark DJ
        • Schnaubelt M
        • Hoti N
        Impact of increased FUT8 expression on the extracellular vesicle proteome in prostate cancer cells.
        J Proteome Res. 2020; 19: 2195-2205
        • Watson JA
        • Burling K
        • Fitzpatrick P
        Urinary insulin-like growth factor 2 identifies the presence of urothelial carcinoma of the bladder.
        BJU Int. 2009; 103: 694-697
        • Kobayashi H
        • Terao T
        • Kawashima Y.
        Serum sialyl Tn as an independent predictor of poor prognosis in patients with epithelial ovarian cancer.
        J Clin Oncol. 1992; 10: 95-101
        • Shimada H
        • Noie T
        • Ohashi M
        • Oba K
        • Takahashi Y.
        Clinical significance of serum tumor markers for gastric cancer: a systematic review of literature by the task force of the Japanese Gastric Cancer Association.
        Gastric Cancer. 2014; 17: 26-33
        • Balog CI
        • Stavenhagen K
        • Fung WL
        N-glycosylation of colorectal cancer tissues: a liquid chromatography and mass spectrometry-based investigation.
        Mol Cell Proteomics. 2012; 11: 571-585
        • Kim GE
        • Bae HI
        • Park HU
        Aberrant expression of MUC5AC and MUC6 gastric mucins and sialyl Tn antigen in intraepithelial neoplasms of the pancreas.
        Gastroenterology. 2002; 123: 1052-1060
        • Molina R
        • Filella X
        • Augé JM
        Tumor markers (CEA, CA 125, CYFRA 21-1, SCC and NSE) in patients with non-small cell lung cancer as an aid in histological diagnosis and prognosis. Comparison with the main clinical and pathological prognostic factors.
        Tumour Biol. 2003; 24: 209-218
        • Vukobrat-Bijedic Z
        • Husic-Selimovic A
        • Sofic A
        Cancer antigens (CEA and CA 19-9) as markers of advanced stage of colorectal carcinoma.
        Med Arch. 2013; 67: 397-401
        • Zhang Z
        • Cheng X
        • Jiang H
        Quantitative proteomic analysis of glycosylated proteins enriched from urine samples with magnetic ConA nanoparticles identifies potential biomarkers for small cell lung cancer.
        J Pharm Biomed Anal. 2021; 206: 1-11
        • Drake PM
        • Cho W
        • Li B
        Sweetening the pot: adding glycosylation to the biomarker discovery equation.
        Clin Chem. 2010; 56: 223-236
        • Pinho SS
        • Reis CA.
        Glycosylation in cancer: mechanisms and clinical implications.
        Nat Rev Cancer. 2015; 15: 540-555
        • Barresi R
        • Campbell KP.
        Dystroglycan: from biosynthesis to pathogenesis of human disease.
        J Cell Sci. 2006; 119: 199-207
        • Yang S
        • Xia J
        • Yang Z
        • Xu M
        • Li S
        Lung cancer molecular mutations and abnormal glycosylation as biomarkers for early diagnosis.
        Cancer Treat Res Commun. 2021; 27: 1-11
        • Nguyen AT
        • Chia J
        • Ros M
        Organelle specific O-glycosylation drives MMP14 activation, tumor growth, and metastasis.
        Cancer Cell. 2017; 32: 639-653
        • Gill DJ
        • Clausen H
        • Bard F.
        Location, location, location: new insights into O-GalNAc protein glycosylation.
        Trends Cell Biol. 2011; 21: 149-158
        • Bennett EP
        • Mandel U
        • Clausen H
        Control of mucin-type O-glycosylation: a classification of the polypeptide GalNAc-transferase gene family.
        Glycobiology. 2012; 22: 736-756
        • Gomes J
        • Mereiter S
        • Magalhães A
        • Reis CA.
        Early GalNAc O-glycosylation: pushing the tumor boundaries.
        Cancer Cell. 2017; 32: 544-545
        • Badhwar A
        • Haqqani AS.
        Biomarker potential of brain-secreted extracellular vesicles in blood in Alzheimer's disease.
        Alzheimers Dement. 2020; 12: 1-14
        • Yang AC
        • Stevens MY
        • Chen MB
        Physiological blood–brain transport is impaired with age by a shift in transcytosis.
        Nature. 2020; 583: 425-430
        • Seol W
        • Kim H
        • Son I.
        Urinary biomarkers for neurodegenerative diseases.
        Exp Neurobiol. 2020; 29: 325-333
        • Fraser KB
        • Moehle MS
        • Alcalay RN
        • West AB.
        Urinary LRRK2 phosphorylation predicts parkinsonian phenotypes in G2019S LRRK2 carriers.
        Neurology. 2016; 86: 994-999
        • Fraser KB
        • Rawlins AB
        • Clark RG
        Ser (P)-1292 LRRK2 in urinary exosomes is elevated in idiopathic Parkinson's disease.
        Mov Disord. 2016; 31: 1543-1550
        • Watanabe Y
        • Hirao Y
        • Kasuga K
        Urinary apolipoprotein C3 is a potential biomarker for Alzheimer's disease.
        Dement Geriatr Cogn Dis Extra. 2020; 10: 94-104
        • Zhang Q
        • Ma C
        • Chin L-S
        • Li L.
        Integrative glycoproteomics reveals protein Nglycosylation aberrations and glycoproteomic network alterations in Alzheimer's disease.
        Sci Adv. 2020; 6: 1-18
        • Chan H-N
        • Xu D
        • Ho S-L
        • Wong MS
        • Li H-W.
        Ultra-sensitive detection of protein biomarkers for diagnosis of Alzheimer's disease.
        Chem Sci. 2017; 8: 4012-4018
        • Disatnik M-H
        • Joshi AU
        • Saw NL
        Potential biomarkers to follow the progression and treatment response of Huntington's disease.
        J Exp Med. 2016; 213: 2655-2669
        • Fujii T
        • Kunugi H.
        p75NTR as a therapeutic target for neuropsychiatric diseases.
        Curr Mol Pharmacol. 2009; 2: 70-76
        • Simmons DA
        • Mills BD
        • Butler III, RR
        Neuroimaging, urinary, and plasma biomarkers of treatment response in Huntington's Disease: preclinical evidence with the p75NTR ligand LM11A-31.
        Neurotherapeutics. 2021; 18: 1039-1063
        • Kim SH
        • Shanware NP
        • Bowler MJ
        • Tibbetts RS.
        Amyotrophic lateral sclerosis-associated proteins TDP-43 and FUS/TLS function in a common biochemical complex to co-regulate HDAC6 mRNA.
        J Biol Chem. 2010; 285: 34097-34105
        • Faruk MO
        • Ichimura Y
        • Kageyama S
        Phase-separated protein droplets of amyotrophic lateral sclerosis-associated p62/SQSTM1 mutants show reduced inner fluidity.
        J Biol Chem. 2021; 297: 1-15
        • Gasparini L
        • Terni B
        • Spillantini MG.
        Frontotemporal dementia with tau pathology.
        Neurodegener Dis. 2007; 4: 236-253
        • Shepheard SR
        • Chataway T
        • Schultz DW
        • Rush RA
        • Rogers M-L.
        The extracellular domain of neurotrophin receptor p75 as a candidate biomarker for amyotrophic lateral sclerosis.
        PLoS One. 2014; 9: 1-9
        • Huo B
        • Zhang W
        • Zhao X
        A triarylphosphine-trimethylpiperidine reagent for the onestep derivatization and enrichment of protein post-translational modifications and identification by mass spectrometry.
        Chem Commun. 2018; 54: 13790-13793
        • Meyer BH
        • Luus HG
        • Muller FO
        Recombinant hirudin, a new anticoagulant, has no effect on faecal blood loss.
        Br J Clin Pharmacol. 1992; 33: 524-526
        • Smet-Nocca C
        • Broncel M
        • Wieruszeski JM
        Identification of O-GlcNAc sites within peptides of the Tau protein and their impact on phosphorylation.
        Mol Biosyst. 2011; 7: 1420-1429
        • Yuzwa SA
        • Yadav AK
        • Skorobogatko Y
        Mapping O-GlcNAc modification sites on tau and generation of a site-specific O-GlcNAc tau antibody.
        Amino Acids. 2011; 40: 857-868
        • Settineri CA
        • Leung I
        • Cousens LS
        Characterization of the glycosylation on recombinant human low-affinity nerve growth factor receptor.
        Techniques in protein chemistry III. Elsevier, 1992: 295-301
        • Rudman N
        • Gornik O
        • Lauc G.
        Altered N-glycosylation profiles as potential biomarkers and drug targets in diabetes.
        FEBS Lett. 2019; 593: 1598-1615
        • Inoue K
        • Wada J
        • Eguchi J
        Urinary fetuin-a is a novel marker for diabetic nephropathy in type 2 diabetes identified by lectin microarray.
        PLoS One. 2013; 8: 1-12
        • Guo Z
        • Liu X
        • Li M
        Differential urinary glycoproteome analysis of type 2 diabetic nephropathy using 2D-LC–MS/MS and iTRAQ quantification.
        J Transl Med. 2015; 13: 1-17
        • Weir GC
        • Ehlers MR
        • Harris KM
        Alpha-1 antitrypsin treatment of new-onset type 1 diabetes: an open-label, phase I clinical trial (RETAIN) to assess safety and pharmacokinetics.
        Pediatr Diabetes. 2018; 19: 945-954
        • Narita T
        • Sasaki H
        • Hosoba M
        Parallel increase in urinary excretion rates of immunoglobulin G, ceruloplasmin, transferrin, and orosomucoid in normoalbuminuric type 2 diabetic patients.
        Diabetes Care. 2004; 27: 1176-1181
        • Golizeh M
        • Lee K
        • Ilchenko S
        Increased serotransferrin and ceruloplasmin turnover in diet-controlled patients with type 2 diabetes.
        Free Radic Biol Med. 2017; 113: 461-469
        • Spiller F
        • Carlos D
        • Souto FO
        α1-Acid glycoprotein decreases neutrophil migration and increases susceptibility to sepsis in diabetic mice.
        Diabetes. 2012; 61: 1584-1591
        • Wang W
        • Xu Y
        • Gao R
        Detection of SARS-CoV-2 in different types of clinical specimens.
        JAMA. 2020; 323: 1843-1844
        • Peng L
        • Liu J
        • Xu W
        SARS-CoV-2 can be detected in urine, blood, anal swabs, and oropharyngeal swabs specimens.
        J Med Virol. 2020; 92: 1676-1680
        • George S
        • Pal AC
        • Gagnon J
        Evidence for SARS-CoV-2 spike protein in the urine of COVID-19 patients.
        Kidney360. 2021; 2: 924-936
        • Watanabe Y
        • Allen JD
        • Wrapp D
        • McLellan JS
        • Crispin M.
        Site-specific glycan analysis of the SARS-CoV-2 spike.
        Science. 2020; 369: 330-333
        • Lau SK
        • Woo PC
        • Wong BH
        Detection of severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein in SARS patients by enzyme-linked immunosorbent assay.
        J Clin Microbiol. 2004; 42: 2884-2889
        • Drosten C
        • Seilmaier M
        • Corman VM
        Clinical features and virological analysis of a case of Middle East respiratory syndrome coronavirus infection.
        Lancet Infect Dis. 2013; 13: 745-751
        • Kirchgesser M
        • Aschenbrenner A
        • Bollwein M
        • Reischl U.
        High-throughput detection of bacterial, fungal and viral nucleic acids in routine microbiological sample types using one generic Pathogen Universal Protocol on the MagNA Pure 96 System.
        MagNA Pure System Application Note No 5 Roche. 2011; : 1-8
        • Tan M
        • Cui L
        • Huo X
        Saliva as a source of reagent to study human susceptibility to avian influenza H7N9 virus infection.
        Emerg Microbes Infect. 2018; 7: 1-10
        • Ping J
        • Lopes TJ
        • Neumann G
        • Kawaoka Y.
        Development of high-yield influenza B virus vaccine viruses.
        Proc Natl Acad Sci USA. 2016; 113: E8296-EE305
        • Nakauchi M
        • Takayama I
        • Takahashi H
        Real-time RT-PCR assays for discriminating influenza B virus Yamagata and Victoria lineages.
        J Virol Methods. 2014; 205: 110-115
        • Canaud G
        • Dejucq-Rainsford N
        • Avettand-Fenoël V
        The kidney as a reservoir for HIV-1 after renal transplantation.
        J Am Soc Nephrol. 2014; 25: 407-419
        • Oelemann WM
        • Lowndes CM
        • Veríssimo da Costa GC
        Diagnostic detection of human immunodeficiency virus type 1 antibodies in urine: a Brazilian study.
        J Clin Microbiol. 2002; 40: 881-885
        • Anyanwu SI
        • Doherty A
        • Powell MD
        Detection of HIV-1 and human proteins in urinary extracellular vesicles from HIV+ patients.
        Adv Virol. 2018; 2018: 1-16
        • Vallari AS
        • Hickman RK
        • Hackett Jr, JR
        Rapid assay for simultaneous detection and differentiation of immunoglobulin G antibodies to human immunodeficiency virus type 1 (HIV-1) group M, HIV-1 group O, and HIV-2.
        J Clin Microbiol. 1998; 36: 3657-3661
        • Ochnio JJ
        • Scheifele DW
        • Ho M
        • Mitchell LA.
        New, ultrasensitive enzyme immunoassay for detecting vaccine-and disease-induced hepatitis A virus-specific immunoglobulin G in saliva.
        J Clin Microbiol. 1997; 35: 98-101
        • Kiyohara T
        • Ouchi Y
        • Hasegawa Y
        An in-house-anti-hepatitis A virus (HAV)specific immunoglobulin M capture enzyme-linked immunosorbent assay: Evaluation and application to an HAV outbreak.
        J Med Virol. 2009; 81: 1513-1516
        • Perry KR
        • Parry JV
        • Vandervelde EM
        Mortimer PP. The detection in urine specimens of IgG and IgM antibodies to hepatitis A and hepatitis B core antigens.
        J Med Virol. 1992; 38: 265-270
        • Wong V
        • Egner W
        • Elsey T
        • Brown D
        • Alexander G.
        Incidence, character and clinical relevance of mixed cryoglobulinaemia in patients with chronic hepatitis C virus infection.
        Clin Exp Immunol. 1996; 104: 25-31
        • Molina A
        • O'Neill MA
        • Darvill AG
        • et al.
        Free glycans as bioactive molecules.
        in: Varki A Cummings RD Esko JD Essentials of glycobiology [Internet]. 4th ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor (NY)2022
        • Hanzawa K
        • Tanaka-Okamoto M
        • Murakami H
        Investigation of acidic free-glycans in urine and their alteration in cancer.
        Glycobiology. 2021; 31: 391-409
        • Ohashi S
        • Iwai K
        • Mega T
        • Hase S.
        Quantitation and isomeric structure analysis of free oligosaccharides present in the cytosol fraction of mouse liver: detection of a free disialobiantennary oligosaccharide and glucosylated oligomannosides.
        J Biochem. 1999; 126: 852-858
        • Xia B
        • Asif G
        • Arthur L
        Oligosaccharide analysis in urine by MALDI-TOF mass spectrometry for the diagnosis of lysosomal storage diseases.
        Clin Chem. 2013; 59: 1357-1368
        • Winchester B.
        Lysosomal metabolism of glycoproteins.
        Glycobiology. 2005; 15: 1R-15R
        • Mak J
        • Cowan TM.
        Detecting lysosomal storage disorders by glycomic profiling using liquid chromatography mass spectrometry.
        Mol Genet Metab. 2021; 134: 43-52
        • Platt FM
        • Boland B
        • van der Spoel AC.
        The cell biology of disease: lysosomal storage disorders: the cellular impact of lysosomal dysfunction.
        J Cell Biol. 2012; 199: 723-734
        • Hanzawa K
        • Tanaka-Okamoto M
        • Murakami H
        Increased levels of acidic free-Nglycans, including multi-antennary and fucosylated structures, in the urine of cancer patients.
        PLoS One. 2022; 17: 1-24
        • Li D
        • Mallory T
        • Satomura S.
        AFP-L3: a new generation of tumor marker for hepatocellular carcinoma.
        Clin Chim Acta. 2001; 313: 15-19
        • Madsen TD
        • Hansen LH
        • Hintze J
        An atlas of O-linked glycosylation on peptide hormones reveals diverse biological roles.
        Nat Commun. 2020; 11: 4033
        • Virreira Winter S
        • Karayel O
        • Strauss MT
        Urinary proteome profiling for stratifying patients with familial Parkinson's disease.
        EMBO Mol Med. 2021; 13: 1-19
        • To KK
        • Lu L
        • Yip CC
        Additional molecular testing of saliva specimens improves the detection of respiratory viruses.
        Emerg Microbes Infect. 2017; 6: 1-7