Prostate cancer (PCa) is the second most common cancer. In this paper, the isolation and properties of exosomes as potential novel liquid biopsy markers for early PCa liquid biopsy diagnosis are investigated using two prostate human cell lines, i.e., benign (control) cell line RWPE1 and carcinoma cell line 22Rv1. Exosomes produced by both cell lines are characterised by various methods including nanoparticle-tracking analysis, dynamic light scattering, scanning electron microscopy and atomic force microscopy. In addition, surface plasmon resonance (SPR) is used to study three different receptors on the exosomal surface (CD63, CD81 and prostate-specific membrane antigen-PMSA), implementing monoclonal antibodies and identifying the type of glycans present on the surface of exosomes using lectins (glycan-recognising proteins). Electrochemical analysis is used to understand the interfacial properties of exosomes. The results indicate that cancerous exosomes are smaller, are produced at higher concentrations, and exhibit more nega tive zeta potential than the control exosomes. The SPR experiments confirm that negatively charged α-2,3- and α-2,6-sialic acid-containing glycans are found in greater abundance on carcinoma exosomes, whereas bisecting and branched glycans are more abundant in the control exosomes. The SPR results also show that a sandwich antibody/exosomes/lectins configuration could be constructed for effective glycoprofiling of exosomes as a novel liquid biopsy marker.
- MeSH
- exozómy * chemie MeSH
- karcinom * metabolismus patologie MeSH
- lektiny analýza metabolismus MeSH
- lidé MeSH
- polysacharidy analýza metabolismus MeSH
- tekutá biopsie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
V práci stručne opisujeme možnosti využitia metód tzv. tekutej biopsie, teda detekcie vysoko nádorovo-špecifických markerov z telových tekutín, na diagnostiku karcinómu prostaty (CaP). V tomto prípade je dôležité hodnotiť biomarkery, ktoré sú orgánovo-špecifické, ako napríklad kalikreíny (rôzne formy PSA), avšak tie nie sú rakovinovo-špecifické. Na zvýšenie spoľahlivosti diagnostiky CaP je preto nutné použiť biomarkery, ktoré sú orgánovo-špecifické a zároveň rakovinovo-špecifické, k takým biomarkerom patrí glykoprofilácia fPSA (free PSA, free prostate-specific antigen/voľný prostatický špecifický antigén) vo forme Giasay (glycopsy immunoassay) testu. Giasay test poskytuje vysokú hodnotu AUC = 0,82 (AUC, area under curve/plocha pod ROC krivkou; ROC, receiver operating curve/operačná charakteristika prijímača), oveľa vyššiu v porovnaní s kalikreínovými testami (vrátane PHI, prostate health index/index zdravia prostaty), a vopred odhalí aj kastračne rezistentnú formu či veľmi skoré štádiá ochorenia. Zároveň Giasay test preukáže podľa výsledkov validácií 63-70 % negatívnych prípadov biopsií v populácii. Tieto hodnoty prevyšujú na Slovensku a v Čechách dostupné laboratórne metódy využívané na skríning a diagnostiku CaP.
In this work, we briefly describe the possibilities of using the so-called liquid biopsy i.e. the detection of highly tumor-specific markers from body fluids, for the diagnosis of prostate cancer (CaP). For this purpose, it is important to use biomarkers that are organ-specific, such as kallikreins (different forms of PSA). Unfortunately, such biomarkers are not cancer-specific. To increase the reliability of CaP diagnosis, it is therefore necessary to use biomarkers that are organ-specific and at the same time cancer-specific, and such biomarkers include glycoprofiling of fPSA (free form of prostate specific antigen) in the form of Giasay (glycopsy immunoassay) test. The Giasay test provides a high value of AUC = 0.82 (AUC - area under curve i.e. the area under receiver operating curve), much higher compared to kallikrein tests (including PHI, prostate health index), and it also reveals the castration-resistant form or very early stages of the disease in advance. At the same time, the Giasay test reveals, according to the validation results, 63-70% of negative cases of biopsies in the population. These values exceed the laboratory methods available in Slovakia and the Czech Republic for the screening and diagnosis of CaP.
- Klíčová slova
- glykoprofilace,
- MeSH
- glykomika * MeSH
- lidé MeSH
- nádorové biomarkery analýza MeSH
- nádory prostaty * diagnóza MeSH
- tekutá biopsie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
BACKGROUND: To compare the clinical performance of a new PCa serum biomarker based on fPSA glycoprofiling to fPSA% and PHI. METHODS: Serum samples from men who underwent prostate biopsy due to increased PSA were used. A comparison between two equal groups (with histologically confirmed PCa or benign, non-cancer condition) was used for the clinical validation of a new glycan-based PCa oncomarker. SPSS and R software packages were used for the multiparametric analyses of the receiver operating curve (ROC) and for genetic algorithm metaheuristics. RESULTS: When comparing the non-cancer and PCa cohorts, the combination of four fPSA glycoforms with two clinical parameters (PGI, prostate glycan index (PGI)) showed an area under receiver operating curve (AUC) value of 0.821 (95% CI 0.754-0.890). AUC values were 0.517 for PSA, 0.683 for fPSA%, and 0.737 for PHI. A glycan analysis was also applied to discriminate low-grade tumors (GS = 6) from significant tumors (GS ≥ 7). CONCLUSIONS: Compared to PSA on its own, or fPSA% and the PHI, PGI showed improved discrimination between presence and absence of PCa and in predicting clinically significant PCa. In addition, the use of PGI would help practitioners avoid 63.5% of unnecessary biopsies, while the use of fPSA% and PHI would help avoid 17.5% and 33.3% of biopsies, respectively, while missing four significant tumors (9.5%).
- Publikační typ
- časopisecké články MeSH
In this paper several advances were implemented for glycoprofiling of prostate specific antigen (PSA), what can be applied for better prostate cancer (PCa) diagnostics in the future: 1) application of Au nanoshells with a magnetic core (MP@silica@Au); 2) use of surface plasmons of Au nanoshells with a magnetic core for spontaneous immobilization of zwitterionic molecules via diazonium salt grafting; 3) a double anti-fouling strategy with integration of zwitterionic molecules on Au surface and on MP@silica@Au particles was implemented to resist non-specific protein binding; 4) application of anti-PSA antibody modified Au nanoshells with a magnetic core for enrichment of PSA from a complex matrix of a human serum; 5) direct incubation of anti-PSA modified MP@silica@Au with affinity bound PSA to the lectin modified electrode surface. The electrochemical impedance spectroscopy (EIS) signal was enhanced 43 times integrating Au nanoshells with a magnetic core compared to the biosensor without them. This proof-of-concept study shows that the biosensor could detect PSA down to 1.2 fM and at the same time to glycoprofile such low PSA concentration using a lectin patterned biosensor device. The biosensor offers a recovery index of 108%, when serum sample was spiked with a physiological concentration of PSA (3.5 ng mL-1).
- MeSH
- biosenzitivní techniky * MeSH
- imobilizační protilátky chemie imunologie MeSH
- impedanční spektroskopie metody MeSH
- lidé MeSH
- nádory prostaty diagnóza patologie MeSH
- nanoslupky chemie MeSH
- prostata patologie MeSH
- prostatický specifický antigen chemie izolace a purifikace MeSH
- zlato chemie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
In recent decades, it has become clear that most of human proteins are glycosylated and that protein glycosylation plays an important role in health and diseases. At present, simple, fast and inexpensive methods are sought for clinical applications and particularly for improved diagnostics of various diseases, including cancer. We propose a label- and reagent-free electrochemical method based on chronopotentiometric stripping (CPS) analysis and a hanging mercury drop electrode for the detection of interaction of sialylated protein biomarker a prostate specific antigen (PSA) with two important lectins: Sambucus nigra agglutinin (SNA) and Maackia amurensis agglutinin (MAA). Incubation of PSA-modified electrode with specific SNA lectin resulted in an increase of CPS peak H of the complex as compared to this peak of individual PSA. By adjusting polarization current and temperature, PSA-MAA interaction can be either eliminated or distinguished from the more abundant PSA-SNA complex. CPS data were in a good agreement with the data obtained by complementary methods, namely surface plasmon resonance and fluorescent lectin microarray. It can be anticipated that CPS will find application in glycomics and proteomics.
Prvé vydanie 169 stran : ilustrace (převážně barevné), portréty ; 24 cm
Kniha poskytuje úvod do problematiky s postupným predstavením tradičných aj moderných biotechnológií, ako aj nanotechnológií a ich praktické aplikácie prevažne v medicíne a molekulárnej biológii. Nakladatelská anotace. Kráceno
- Konspekt
- Biotechnologie. Genetické inženýrství
- NLK Obory
- biomedicínské inženýrství
- NLK Publikační typ
- kolektivní monografie
An ultrasensitive impedimetric glycan-based biosensor for reliable and selective detection of inactivated, but intact influenza viruses H3N2 was developed. Such glycan-based approach has a distinct advantage over antibody-based detection of influenza viruses since glycans are natural viral receptors with a possibility to selectively distinguish between potentially pathogenic influenza subtypes by the glycan-based biosensors. Build-up of the biosensor was carefully optimized with atomic force microscopy applied for visualization of the biosensor surface after binding of viruses with the topology of an individual viral particle H3N2 analyzed. The glycan biosensor could detect a glycan binding lectin with a limit of detection (LOD) of 5 aM. The biosensor was finally applied for analysis of influenza viruses H3N2 with LOD of 13 viral particles in 1 μl, what is the lowest LOD for analysis of influenza viral particles by the glycan-based device achieved so far. The biosensor could detect H3N2 viruses selectively with a sensitivity ratio of 30 over influenza viruses H7N7. The impedimetric biosensor presented here is the most sensitive glycan-based device for detection of influenza viruses and among the most sensitive antibody or aptamer based biosensor devices.
- MeSH
- biosenzitivní techniky metody MeSH
- chřipka lidská diagnóza virologie MeSH
- impedanční spektroskopie metody MeSH
- lidé MeSH
- limita detekce MeSH
- polysacharidy chemie MeSH
- virus chřipky A, podtyp H3N2 izolace a purifikace MeSH
- virus chřipky A, podtyp H7N7 izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- elektrochemické techniky přístrojové vybavení metody MeSH
- glykomika přístrojové vybavení metody MeSH
- glykoproteiny analýza metabolismus MeSH
- lidé MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- proteiny analýza metabolismus MeSH
- sacharidové sekvence MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Glycan biochips and biosensors are potentially important tools for detection of glycan – protein interactions. Among several applications they can be used for rapid and precise diagnosis of various diseases and infections. Two major detection techniques available for construction of glycan biochips and biosensors – fluorescent labelled and label-free methods are discussed.
- MeSH
- bakteriální infekce * diagnóza MeSH
- biosenzitivní techniky * metody přístrojové vybavení využití MeSH
- chemické techniky analytické MeSH
- diagnostické zobrazování metody využití MeSH
- DNA sondy * MeSH
- fluoroimunoanalýza metody využití MeSH
- lidé MeSH
- optické zobrazování metody využití MeSH
- polysacharidy analýza chemie MeSH
- povrchová plasmonová rezonance metody přístrojové vybavení využití MeSH
- proteiny chemie MeSH
- spektrální analýza metody využití MeSH
- virové nemoci * diagnóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
