Despite numerous studies, isolating pure preparations of extracellular vesicles (EVs) has proven challenging. Here, we compared ion-exchange chromatography (IEC) to the widely used sucrose density gradient (SDG) centrifugation method for purification EVs. EVs in bulk were isolated from pooled normal human amniotic fluid (AF) by differential followed IEC or separation. The purity was evaluated electrophoresis and lectin blotting/immuno blotting monitor distribution total proteins, different...

Urine is a readily available source of relatively large quantities extracellular vesicles (EVs). However, the isolation urinary EVs (uEVs) complicated by presence Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as contaminant. This may make glycan analysis uEVs difficult since THP heavily glycosylated. To facilitate glycosylation address need for elimination non-uEV glycans, we present modification uEV procedure use isolated in development lectin-exosome binding assay....

The beta-galactoside-binding proteins galectin-1 and -3 are thought to modulate cell-extracellular matrix interactions in cell adhesion migration. In this study, their occurrence human trophoblast has been investigated. the first trimester placenta is expressed cytotrophoblast of mid distal columns, but absent from villous proximal column cytotrophoblast. syncytiotrophoblast was also positive. Galectin-3, on other hand, uniformly localized columns. Immunolocalization these placental bed...

CA125, a coelomic epithelium-related antigen, is expressed in both normal and pathological conditions. In this study, we compared the glycosylation of CA125 antigen from amniotic fluid ovarian carcinoma cell line OVCAR-3, order to detect p

Glycans as Biomarkers: Status and Perspectives Protein glycosylation is a ubiquitous complex co- post-translational modification leading to glycan formation, i.e. oligosaccharide chains covalently attached peptide backbones. The significance of changes in for the beginning, progress outcome different human diseases widely recognized. Thus, glycans are considered unique structures diagnose, predict susceptibility monitor progression disease. In »omics« era, glycome, analogue proteome genome,...

This study was aimed at obtaining insight into the diversity of sialic acids in cancer- and non-cancer-related CA125 antigen, tumour marker serous ovarian cancer. Starting from available data suggesting possible relevance for discriminating antigens different origin, we have employed a new experimental approach based on use human acid-binding Ig-like lectins, Siglecs, as tools investigation sialylation. Siglec-2, belonging to group evolutionarily conserved Siglec-3, -6, -7, -9 -10, which are...

This study was aimed at obtaining insight into the diversity of sialic acids in cancer- and non-cancer-related CA125 antigen, tumour marker serous ovarian cancer. Starting from available data suggesting possible relevance for discriminating antigens different origin, we have employed a new experimental approach based on use human acid-binding Ig-like lectins, Siglecs, as tools investigation sialylation. Siglec−2, belonging to group evolutionarily conserved Siglec−3, −6, −7, −9 −10, which are...

The cancer antigen CA125 has a very complex molecular architecture in terms of both protein backbone and oligosaccharide chains. In this study, we examined the forms microheterogeneity chains pregnancy-associated CA125, as first step towards gaining an insight into its possible involvement ligand carbohydrate-dependent interactions. glycobiochemical properties may be diagnostic biomedical importance specific markers physiological pathological conditions early pregnancy, well targets...

Fibronectin (FN) is a multifunctional glycoprotein involved in cell‐matrix interactions. It exhibits complex pattern of forms differing respect to aminoacid and oligosaccharide composition. In this study we examined glycobiochemical functional properties the FN benign prostatic hyperplasia (BPH) cancer (PCa), attempting resolve disease‐related differences. Two BPH sera pools three PCa were used as source. The affinity‐purified molecule was characterized by SDS‐PAGE, immuno‐ lectin blot,...

Background: Extracellular vesicles (EVs), released from the plasma membrane or intracellular compartments, have a specific composition related to their parent cells, but they can, additionally, be modified by extracellular environment. Although glycans are known contribute EV and may biomedical importance as biomarkers recognition signals, not been extensively investigated. In this study, seminal prostasomes, i.e. EVs (SP) of normo- oligozoospermic men, were analyzed in order detect possible...

CA125 antigen is a mucin-type molecule with complex protein backbone and oligosaccharide chain structure. In this study, we characterized from human amniotic fluid by gel filtration, ion-exchange chromatography, peptide mass fingerprinting lectin-binding assays. The obtained results indicate to be structurally heterogeneous, existing in different glycoisoforms subtle differences the profile of molecular forms comparison placental tissue-derived cancer-derived antigen. complexity structure...

Defining the molecular characteristics of seminal plasma proteins is essential for understanding their function in physiological and pathological conditions.Starting from predicted importance human gelatin-binding proteins, comprising fibronectin (FN) FN-related molecules, male fertility, this study aims at gaining insight into immuno-glycobiochemical properties.Human subjects with normal semen parameters were separated on a gelatin-Sepharose column analyzed by sodium dodecyl...

CA125 antigen is a high molecular mass, structurally heterogeneous, mucin-type molecule expressed during embryonic development as well in adult human tissues. This study was aimed at investigating its mucin-related property of ferning, general complementary way characterization. Pregnancy-associated (pCA125) examined using light, transmission and scanning electron microscopy compared with cancer-derived (cCA125). The results obtained for spread-out, air-dried pCA125 cCA125 samples revealed...

Background: Human seminal prostasomes are intrinsically heterogeneous extracellular vesicles (EVs) whose composition is, additionally, influenced by different physiological conditions. Aiming at the molecular properties of prostasomal surface exemplified glycan compositions as a possible distinction factor, we applied lectin-affinity chromatography (LAC) new tool for their separation. Since glycans, generally, exhibit various biological activities, introduction glyco-parameters reference...

This study was aimed at defining molecular species of prostate-specific antigen (PSA) in immune complexes with immunoglobulin M (IgM). Having mind the oligoreactivity IgM and its preference for carbohydrate antigens, there is possibility that it can selectively recognize known PSA glycoisoforms. PSA-IgM free fractions were separated from sera subjects prostate cancer (PCa) benign prostatic hyperplasia (BPH) by gel filtration subjected to on-chip immunoaffinity ion-exchange chromatography....

High levels of expression galectin-1 and galectin-3, the β-galactoside-binding proteins, have been recently described in malignant thyroid tumors but not adenomas nor normal tissue. However, there are no data about these galectins during fetal development. In this study we analyzed immunohistochemically presence galectin-3 human glands (16–37 weeks gestation). Weak to moderate cytoplasmic staining for was observed follicular cells all thyroids. Galectin-3 could be detected any investigated....

Glycans as a Target in the Detection of Reproductive Tract Cancers The significance changes glycosylation for beginning, progress and outcome different human diseases is highly recognized. In this review we summarized literature data on alteration glycans cancer, especially glycoforms tumor markers reproductive tract cancers: prostate-specific antigen (PSA) cancer 125 (CA125). We aimed to highlight diagnostic potential relevance glycan microheterogeneity present some novel methods detection....

A soluble carbohydrate-binding protein with galactoside-binding properties (CBP14) was isolated from liver nuclei of adult rats. Proteins were extracted membrane-depleted nuclei; they contained 3 major fractions (Mr 43,000, 14,000 and 10,000) which reacted antibodies raised against human placental β-galactoside-binding lectin (HP 14) as well asialofetuin (ASF). Enrichment (72- to 143-fold) CBP14 carried out by affinity chromatography, using anti-HP 14 ASF consequent ligands. retained column...

Prostate specific antigen (PSA) exhibits pronounced heterogeneity in both primary structure and glycan composition, resulting the existence of different molecular forms. Investigation PSA is a demanding task facing limitations due to inadequate sensitivity analytical techniques low concentrations This study aimed profile free (fPSA), especially lower mass species lacking detailed classification, normal seminal plasma sera from subjects with benign hyperplasia (BPH) or cancer prostate (PCa)...