کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
866235 | 1470948 | 2016 | 7 صفحه PDF | دانلود رایگان |

• The sensitive detection of HIF1α achieved through the antibody-antigen immune complex formation.
• HIF1α was detected electrochemically in MCF-7 cells and distinguished it from normal cells.
• The expression of HIF1α was compared between 3 different cancer cell lines MCF-7, PC-3, and A549.
• Hypoxia was induced chemically using hypoxic mimicking agents and the effect was examined in A549 cell lines.
Hypoxia inducible factor 1 alpha (HIF1α) overexpression was detected in cancerous cells using an amperometric immunosensor with a nano-bioconjugate. The sensor probe was fabricated by covalently immobilizing the antibody (anti-HIF1α) onto a composite layer of functionalized conducting polymer [2,2:5,2-terthiophene-3-(p-benzoic acid)] (pTTBA) formed on a layer of gold nanoparticles (AuNPs). A nano-bioconjugate with hydrazine and a secondary antibody of HIF1α (sec-Ab2) attached on AuNPs reveals the immunoreaction at the sensor probe through the catalytic reduction of H2O2 by hydrazine at −0.35 V vs. Ag/AgCl. Morphology and performance of the sensor probe were characterized using FE-SEM, XPS, EIS, and cyclic voltammetry. The calibration plot at optimized experimental conditions shows a dynamic range of 25–350 pM/mL with a detection limit of 5.35±0.02 pM/mL. The reliability of the sensor was evaluated using non-cancerous Vero and cancerous MCF-7 cell lysates, where the HIF1α expression was compared with three cancerous cell lines MCF-7, PC-3, and A549. Furthermore, the sensor probe confirms the stable expression of HIF1α in the A549 lung cancer cells when exposing them to hypoxic mimicking agents Co, Ni, and Mn ions. Of these, Co ions show the highest stabilization effect on HIF1α followed by Ni and Mn ions, respectively.
Journal: Biosensors and Bioelectronics - Volume 83, 15 September 2016, Pages 312–318