HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES
Year 2021,
Volume: 22 Issue: 4, 344 - 352, 29.12.2021
Fatma Doğan Guzel
,
Iremnur Akcakoca
Hamed Ghorbanpoor
,
Araz Norouz Dizaji
Yasin Öztürk
Ewen Blair
Tanıl Kocagoz
Damion Corrigan
Hüseyin Avcı
Abstract
Heat shock proteins are produced when the organisms are exposed to various environmental stress conditions such as temperature, light, toxins. It is a known fact that in bacteria, which has the HSP gene, antibiotics can trigger the expression of the heat shock protein. However, the response of heat shock protein genes to antibiotics has not been fully clarified in the literature yet, studies are still ongoing. In this study, a novel way for the detection heat shock protein65 was investigated using the electrochemical impedance spectroscopy due to its sensitivity, selectivity, low cost. To do so, heat shock protein65 probe and target were designed and the hybridization behavior of the probe with designed target was studied upon the binding onto screen-printed electrodes. Cyclic voltammetry was performed to analyze surface characterization of secreen printed electrodes and the performance of the screen printed electrodes was tested using electrocehimcal impedance spectroscopy by measuring changes in the charge transfer resistance upon hybridization. Based on obtained results, the designed heat shock protein65 probe was confirmed and an appropriate increase in charge transfer resistance values compliance to the literature proved that the electrochemical impedance spectroscopy can be effectively used to detect heast shock protein65 probe label-free. Results presented here can lead to development of antibiotic susceptibility assay based on the heat shock protein genes in future.
Supporting Institution
TÜBİTAK
Thanks
This study was conducted in the frame of Newton Katip Celebi Fund between Turkey and UK and supported by Turkish Scientific and Technological Council (TUBITAK) under the grant number of 217S793. Authors would like to thank I. A. Ince for useful discussions.
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Year 2021,
Volume: 22 Issue: 4, 344 - 352, 29.12.2021
Fatma Doğan Guzel
,
Iremnur Akcakoca
Hamed Ghorbanpoor
,
Araz Norouz Dizaji
Yasin Öztürk
Ewen Blair
Tanıl Kocagoz
Damion Corrigan
Hüseyin Avcı
References
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- Beere HM. “The stress of dying”: The role of heat shock proteins in the regulation of apoptosis. J Cell Sci 2004; 117: 2641–2651.
- Nguyen L. Antibiotic resistance mechanisms in M. tuberculosis: an update. Arch Toxicol 2016; 90: 585–1604.
- Hendrick JP, Hartl F. Functıons of Heat-Shock 1993.
- Tran TD, Kwon HY, Kim EH, Kim KW, Briles DE, Pyo S, Rhee DK. Decrease in penicillin susceptibility due to heat shock protein ClpL in Streptococcus pneumoniae. Antimicrob Agents Chemother 2011; 55: 2714–2728.
- Briffotaux J, Liu S, Gicquel B. Genome-wide transcriptional responses of Mycobacterium to antibiotics. Front Microbiol 2019; 10: 1–14.
- A. Mudaliar AV, Kashyap RS, Purohit HJ, Taori GM, Daginawala HF. Detection of 65 kD heat shock protein in cerebrospinal fluid of tuberculous meningitis patients. BMC Neurol 2006; 6: 1–7.
- Senna SG, Battilana J, Costa JC, Silva MG, Duarte RS, Fonseca LS, Suffys PN, Bogo MR. Sequencing of hsp65 gene for identification of Mycobacterium species isolated from environmental and clinical sources in Rio de Janeiro, Brazil. J Clin Microbiol 2008; 46: 3822–3825.
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- Onat Akbulut S, Ghorbanpoor H, İpteç BÖ, Butterworth A, Avcıoğlu G, Kozacı LD, Topateş G, Corrigan DK, Avci H, Guzel FD. Impedance testing of porous Si3N4 scaffolds for skeletal implant applications SN Appl Sci 2020; 2: 823-828.
- Guzel FD, Citak F. Development of an on-chip antibiotic permeability assay with single molecule detection capability. IEEE Trans Nanobioscience 2018; 17: 155–160.
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- Baselt DR, Lee GU, Natesan M, Metzger SW, Sheehan PE, Colton RJ. A biosensor based on magnetoresistance technology. Biosens Bioelectron 1998; 13: 731–739.
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- Blair EO, Corrigan DK. A review of microfabricated electrochemical biosensors for DNA detection. Biosens Bioelectron 2019; 134: 57–67.
- Liu J, Cao Z, Lu Y. Functional nucleic acid sensors. Chem Rev 2009; 109: 1948–1998.
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- Butterworth A, Blues E, Williamson P, Cardona M, Gray L, Corrigan DK. SAM composition and electrode roughness affect performance of a DNA biosensor for antibiotic resistance. Biosens 2019; 9: 1-12.
- Bedford E. Gold surface nanostructuring for separation and sensing of biomolecules. Doctoral dissertation 2017.
- Keighley SD, Li P, Estrela P, Migliorato P. Optimization of DNA immobilization on gold electrodes for label-free detection by electrochemical impedance spectroscopy. Biosens Bioelectronvol 2008; 23: 1291–1297.
- Guzel FD, Ghorbanpoor H, Dizaji AN, Akcakoca I, Ozturk Y, Kocagoz T, Corrigan DK, Avci H. Biotechnol Appl Biochem 2020; doi:10.1002/bab.2037.
- Lisdat F, Schäfer D. The use of electrochemical impedance spectroscopy for biosensing. Anal Bioanal Chem 2008; 391: 1555–1567.
- Park JY, Park SM. DNA hybridization sensors based on electrochemical impedance spectroscopy as a detection tool. Sensors 2009; 9: 9513–9532.
- Drummond TG, Hill MG, Barton JK. Electrochemical DNA sensors. Nat Biotechnol 2003; l: 1192–1199.
- Demirbakan B, Sezgintürk MK. A novel immunosensor based on fullerene C60 for electrochemical analysis of heat shock protein 70. J Electroanal Chem 2016; 783: 201–207.
- Sun B, Cai J, Li W, Gou X, Gou Y, Li D, Hu F. A novel electrochemical immunosensor based on PG for early screening of depression markers-heat shock protein 70. Biosens Bioelectron 2018; 111: 34–40.
- Aguilar ZP, Fritsch I. Immobilized Enzyme-Linked DNA-Hybridization Assay with Electrochemical Detection for Cryptosporidium p arvum hsp70 mRNA. Anal Chem 2003; 75: 3890–3897.