2015
3D-printed biosensor with poly(dimethylsiloxane) reservoir for magnetic separation and quantum dots-based immunolabeling of metallothionein
HEGER, Zbynek; Jan ZITKA; Natalia CERNEI; Sona KRIZKOVA; Markéta SZTALMACHOVÁ et al.Základní údaje
Originální název
3D-printed biosensor with poly(dimethylsiloxane) reservoir for magnetic separation and quantum dots-based immunolabeling of metallothionein
Autoři
HEGER, Zbynek; Jan ZITKA; Natalia CERNEI; Sona KRIZKOVA; Markéta SZTALMACHOVÁ; Pavel KOPEL; Michal MASAŘÍK; Petr HODEK; Ondrej ZÍTKA; Vojtech ADAM a Rene KIZEK
Vydání
Electrophoresis, Hoboken, Wiley-Blackwell, 2015, 0173-0835
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 2.482
Označené pro přenos do RIV
Ano
Kód RIV
RIV/00216224:14110/15:00087449
Organizační jednotka
Lékařská fakulta
UT WoS
EID Scopus
Klíčová slova anglicky
Biosensor; Bioseparation; Head and neck cancer; Metallothionein; Nanotechnology
Štítky
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 22. 9. 2015 15:15, Soňa Böhmová
Anotace
V originále
Currently, metallothioneins (MTs) are extensively investigated as the molecular biomarkers and the significant positive association of the MT amount was observed in tumorous versus healthy tissue of various types of malignant tumors, including head and neck cancer. Thus, we proposed a biosensor with fluorescence detection, comprising paramagnetic nanoparticles (nanomaghemite core with gold nanoparticles containing shell) for the magnetic separation of MT, based on affinity of its sulfhydryl groups toward gold. Biosensor was crafted from PDMS combined with technology of 3D printing and contained reservoir with volume of 50 mu L linked to input (sample/detection components and washing/immunobuffer) and output (waste). For the immunolabeling of immobilized MT anti-MT antibodies conjugated to CdTe quantum dots through synthetic heptapeptide were employed. After optimization of fundamental conditions of the immunolabeling (120 min, 20 degrees C, and 1250 rpm) we performed it on a surface of paramagnetic nanoparticles in the biosensor reservoir, with evaluation of fluorescence of quantum dots (lambda(exc) 400 nm, and lambda(em) 555 nm). The developed biosensor was applied for quantification of MT in cell lines derived from spinocellular carcinoma (cell line 122P-N) and fibroblasts (122P-F) and levels of the biomarker were found to be about 90 nM in tumor cells and 37 nM in fibroblasts. The proposed system is able to work with low volumes (< 100 mu L), with low acquisition costs and high portability.
Návaznosti
| NT14337, projekt VaV |
|