J 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.

Basic information

Original name

3D-printed biosensor with poly(dimethylsiloxane) reservoir for magnetic separation and quantum dots-based immunolabeling of metallothionein

Authors

HEGER, Zbynek (203 Czech Republic), Jan ZITKA (203 Czech Republic), Natalia CERNEI (203 Czech Republic), Sona KRIZKOVA (203 Czech Republic), Markéta SZTALMACHOVÁ (203 Czech Republic, belonging to the institution), Pavel KOPEL (203 Czech Republic), Michal MASAŘÍK (203 Czech Republic, guarantor, belonging to the institution), Petr HODEK (203 Czech Republic), Ondrej ZÍTKA (203 Czech Republic), Vojtech ADAM (203 Czech Republic) and Rene KIZEK (203 Czech Republic)

Edition

Electrophoresis, Hoboken, Wiley-Blackwell, 2015, 0173-0835

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 2.482

RIV identification code

RIV/00216224:14110/15:00087449

Organization unit

Faculty of Medicine

UT WoS

000356004200005

Keywords in English

Biosensor; Bioseparation; Head and neck cancer; Metallothionein; Nanotechnology

Tags

Tags

International impact, Reviewed
Změněno: 22/9/2015 15:15, Soňa Böhmová

Abstract

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.

Links

NT14337, research and development project
Name: Studium a charakterizace primárních nádorových buněčných linií spinocelulárních karcinomů v oblasti hlavy a krku a jejich maligní potenciál.
Investor: Ministry of Health of the CR