Table 2.
Serum glycoproteins with altered glycosylation identified in pancreatic cancer patients
| Techniques | Serum cohort (n) | Serum depletion (Yes/No) | Glycosylation change | Main glycoproteins identified | Ref. |
| ConA chromatography. Tryptic digestion. N-glycan MS analysis of the N-glycopeptides | 2 PaC | No | Increased branching, fucosylation and sialylation | [101] | |
| ConA and WGA chromatography. Glycoprotein separation and array with lectins (SNA, PNA, MAL, AAL). | 6 PaC 8 ChrP 10 HC | Yes (IgY 12) (albumin, IgG and major acute-phase proteins) | Increased fucosylation and sialylation | Hemopexin Serum amyloid P component Antithrombin-III Haptoglobin (HPT)-related protein β-2-glycoprotein 1 | [110] |
| Antibody microarray to α1-β-glycoprotein, amyloid P and antithrombin followed by lectin detection (SNA). MS glycoprotein identification. | 22 PaC (III/IV) 35 ChrP 89 HC 37 diabetic | Increase affinity to SNA (α2,6-sialic acid) by 69% in PaC patients | α1-β-glycoprotein. ROC curve (cancer vs non cancer samples), 100% sensitivity 98% specificity. AUC = 0.998 | [111] | |
| Bead-based antibody-lectin (SNA, Con A) multiplex assay-for determining SNA and Con A reactivity of α1-β-glycoprotein, and amyloid P component. | 20 PaC (III/IV) 20 ChrP 20 HC | SNA affinity (α2,6-sialic acid) | α2,6-sialic glycoforms of α1-β-glycoprotein Differentiation of ChrP vs PaC (P = 0.035) | [112] | |
| SNA affinity chromatography to enrich sialylated glycopeptides and compared their relative abundance by ultra performance LC-MS | 10 PaC (II-III) 5 Acute Pancreatitis 16 HC | Yes (albumin depleted) | SNA (α2,6-sialic acid) | Sialylated glycopeptides of HPT, α-1-antitrypsin (A1AT), transferrin, ceruloplasmin, α1-acid-glycoprotein (AGP), fetuin A and Igs. Change in acute pancreatitis and PaC | [97] |
| 2DE followed by N-glycan sequencing | 9 PaC (I-IV) 3 ChrP 5HC | SLex Fucosylation | Increase in SLex on AGP, HPT and transferrin in advanced PaC and ChrP Increase in core fucosylation of HPT and AGP in PaC vs ChrP and HC | [113] | |
| Electrophoresis (1DE) followed by WB with anti SLex. Immunoprecipitation of ceruloplasmin and SLex detection | 20 PaC (IIa-IV) 14 ChrP 13 HC | Yes (IgY 12) (albumin, IgG and major acute-phase proteins | SLex | Ceruloplasmin Tendency to an increase of SLex on ceruloplasmin in PaC vs HC and ChrP | [114] |
| Lectin (AAL)-antibody ELISA | 72 PaC 22 HC 63 pancreatitis | AAL (fucosylation). | Increase of fucosylated HPT in advanced PaC | [115] | |
| AGP purification MS analysis of AGP N-glycans and AAL ELISA | 19 PaC (I-IV) 6 ChrP 6 HC | α1,3 fucosylation | Increase of fucosylated AGP in advanced PaC | [116-118] | |
| N-glycan sequencing of human serum ribonuclease (RNase 1). | 2 PaC 2 HC | Core fucosylation | Increase of core fucosylation in RNase 1 in PaC | [120] | |
| ELISA to measure N-glycosylation Asn-88 site occupancy of serum RNase 1 | 91 PaC 60 HC | Asn-88 N-glycosylation | Increase in N-glycosylated Asn-88 of RNase 1 (normalized to RNase 1) in PaC. | [121] | |
| AAL to enrich fucosylated glycoproteins LC-MS/MS analyses ELISA/lectin ELISAs | 20 IPMN 10 MCN 37 PaC (I-IV) 30 HC 30 ChrP 22 OJ 30 Type II DM | IgY-14 LC10 columns | Fucosylation (AAL) | ACT trombospondin-1 HPT High diagnostic potential combined with CA 19-9 | [119] |
| nanoLC-MS/MS analysis of iTRAQ labelled glycopeptides. | 13 HC 13 ChrP 13 PaC 1 Std | IgY-14 LC10 column | Core-fucosylation | One core fucosylated peptide from ACT different between groups | [108] |
| PHA-L lectin to enrich complex N-glycoproteins 2D nanoLC-MS/MS analyses Western Blot with biotinylated PHA-L nanoLC-MS/MS of tryptic digested gel bands that corresponded to specific lectin interactions on Western blot | 26 HC (include ChrP + pseudo cysts) 76 PaC | Albumin/IgG depletion | Increased fucosylation N211 Novel glycosylation site N64 New N-glycosylated side at N2336 in PaC N-glycosylation N877 | HPT Leukemia inhibitory factor receptor LIFR Centrosome-associate protein 350 CE350 Vacuolar protein sorting-associated protein 13A VP13A | [122] |
| 2D-LC-MS/MS | 231 serum women samples pooled in groups: time-to-diagnosis | Yes | N-glycosylation occupancy | A1AT HPT AGP | [123] |
| Lectin (CCL2)-antibody ELISA | 109 PaC 91 control (plasma) | No | CCL2 (3’ fucosylation) | MUC5AC | [99] |
| Antibody lectin sandwitch array | 156 PaC (I-IV) 160 control (plasma) | No | SLea related | MUC5AC MUC16 | [127] |
| Antibody microarray capture of proteins. Glycan analysis with lectins (AAL, WGA) and CA 19-9 | 23 PaC (I-IV) 23 HC | No | CA 19-9 | MUC1 CEA | [128] |
| Antibody lectin sandwich array | 23 PaC (I-IV) 23 HC | No | SLea(CA 19-9) | MUC1 MUC5AC | [129] |
| Antibody array | 285 PaC (I-IV) 102 ChrP 144 HC (serum & plasma) | No | CA 19-9 | MUC1 MUC5AC MUC16 | [130] |
MS: Mass-spectrometry; PaC: Pancreatic cancer; WGA: Wheat germ agglutinin; SNA: Sambucus nigra agglutinin; AAL: Aleuria aurantia lectin; ChrP: Chronic pancreatitis; HC: Healthy controls.