Nejvíce citovaný článek - PubMed ID 25829219
Rapid and sensitive detection of multiple microRNAs in cell lysate by low-fouling surface plasmon resonance biosensor
Cells are continuously sensing their microenvironment and subsequently respond to different physicochemical cues by the activation or inhibition of different signaling pathways. To study a very complex cellular response, it is necessary to diminish background environmental influences and highlight the particular event. However, surface-driven nonspecific interactions of the abundant biomolecules from the environment influence the targeted cell response significantly. Yes-associated protein (YAP) translocation may serve as a marker of human hepatocellular carcinoma (Huh7) cell responses to the extracellular matrix and surface-mediated stresses. Here, we propose a platform of tunable functionable antifouling poly(carboxybetain) (pCB)-based brushes to achieve a molecularly clean background for studying arginine, glycine, and aspartic acid (RGD)-induced YAP-connected mechanotransduction. Using two different sets of RGD-functionalized zwitterionic antifouling coatings with varying compositions of the antifouling layer, a clear correlation of YAP distribution with RGD functionalization concentrations was observed. On the other hand, commonly used surface passivation by the oligo(ethylene glycol)-based self-assembled monolayer (SAM) shows no potential to induce dependency of the YAP distribution on RGD concentrations. The results indicate that the antifouling background is a crucial component of surface-based cellular response studies, and pCB-based zwitterionic antifouling brush architectures may serve as a potential next-generation easily functionable surface platform for the monitoring and quantification of cellular processes.
- Klíčová slova
- antifouling polymer brushes, cell mechanotransduction, cell signaling, functional biointerfaces, surface modification, zwitterionic material,
- MeSH
- akrylamidy chemie MeSH
- biokompatibilní potahované materiály chemie MeSH
- bioznečištění prevence a kontrola MeSH
- buněčný převod mechanických signálů * MeSH
- extracelulární matrix metabolismus MeSH
- lidé MeSH
- mechanický stres MeSH
- nádorové buněčné linie MeSH
- oligopeptidy chemie MeSH
- protoonkogenní proteiny c-yes metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylamidy MeSH
- arginyl-glycyl-aspartic acid MeSH Prohlížeč
- biokompatibilní potahované materiály MeSH
- oligopeptidy MeSH
- protoonkogenní proteiny c-yes MeSH
- YES1 protein, human MeSH Prohlížeč
- zwitterion carboxybetaine acrylamide MeSH Prohlížeč
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematological malignancies with a high risk of transformation to acute myeloid leukemia (AML). MDS are associated with posttranslational modifications of proteins and variations in the protein expression levels. In this work, we present a novel interactomic diagnostic method based on both protein array and surface plasmon resonance biosensor technology, which enables monitoring of protein-protein interactions in a label-free manner. In contrast to conventional methods based on the detection of individual biomarkers, our presented method relies on measuring interactions between arrays of selected proteins and patient plasma. We apply this method to plasma samples obtained from MDS and AML patients, as well as healthy donors, and demonstrate that even a small protein array comprising six selected proteins allows the method to discriminate among different MDS subtypes and healthy donors.
- MeSH
- analýza hlavních komponent MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mapování interakce mezi proteiny * MeSH
- mladý dospělý MeSH
- myelodysplastické syndromy krev diagnóza MeSH
- povrchová plasmonová rezonance MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- vazba proteinů MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH