HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT
SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES

DIZAJI, Araz Norouz; AKCAKOCA, Iremnur; BLAIR, Ewen O.; OZTURK, Yasin; CORRIGAN, Damion K; GHORBANPOOR, Hamed; GUZEL, Fatma DOGAN; KOCAGOZ, Tanil

doi:10.18038/estubtda.806222

HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES

Fatma DOGAN GUZEL, (Biyomedikal Mühendisliği Bölümü, Mühendislik ve Doğa Bilimleri Fakültesi, Ankara Yıldırım Beyazıt Üniversitesi, Ankara Türkiye)

Iremnur AKCAKOCA, (Mühendislik ve Doğa Bilimleri Fakültesi Metalurji ve Malzeme Mühendisliği Bölümü Yıldırım Beyazıt Üniversite, Ankara, Türkiye)

Hamed GHORBANPOOR, (Biyomedikal Mühendisliği Bölümü, Eskişehir Osmangazi Üniversitesi, Eskişehir, Türkiye)

Araz Norouz DIZAJI, (Biyomedikal Mühendisliği Bölümü, Mühendislik ve Doğa Bilimleri Fakültesi, Ankara Yıldırım Beyazıt Üniversitesi, Ankara Türkiye)

Yasin OZTURK, (Mühendislik ve Doğa Bilimleri Fakültesi Metalurji ve Malzeme Mühendisliği Bölümü Yıldırım Beyazıt Üniversite, Ankara, Türkiye)

Ewen O. BLAIR, (Biyomedikal Mühendisliği Bölümü, Strathclyde Üniversitesi, G1 1XQ, Birleşik Krallık)

Tanil KOCAGOZ, (Sağlık Bilimleri Enstitüsü, Tıbbi Biyoteknoloji Anabilim Dalı, İstanbul, Türkiye)

Damion K CORRIGAN (Biyomedikal Mühendisliği Bölümü, Strathclyde Üniversitesi, G1 1XQ, Birleşik Krallık)

Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering

2 0

Yıl: 2021 Cilt: 22 Sayı: 4 Sayfa Aralığı: 344 - 352 Metin Dili: İngilizce DOI: 10.18038/estubtda.806222 İndeks Tarihi: 23-06-2022

HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES

Öz:

Heat shock proteins (HSPs) are produced when organisms are exposed to various environmental stress conditions such as extreme temperatures, light, and toxins. It is a known fact that in bacteria which have the HSP gene (hsp), antibiotics can trigger the expression of these proteins. However, the response of HSP genes to antibiotics has not been fully clarified in the literature, with studies still ongoing. In this work, a novel method of detecting heat shock protein 65 (HSP65) was investigated using electrochemical impedance spectroscopy (EIS) due to its sensitivity and selectivity. To do so, a specific HSP65 probe and target were designed and their hybridization behaviour was studied using low-cost screen-printed electrodes (SPEs). Cyclic voltammetry was performed to analyse surface characteristics of the SPEs and the performance of the electrodes was tested using EIS, by measuring changes in the charge transfer resistance upon probe binding and target hybridization. Increases in charge transfer resistance were measured and observed to be in-line with literature. Based on these results, the designed HSP65 probe was confirmed to bind the target sequence and proved that the EIS can be effectively used to detect HSP65 label-free. Results presented here could lead to development of antibiotic susceptibility assays based on hsp genes in future and provide a quicker test for the detection of many slow-growing bacteria

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

[1] Garred P, Brygge K, Sorensen CH, Madsen HO, Thiel S, Svejgaard A. Mycobacteria in water. J Appl Bacteriol 1984; 57: 193–211.
[2] Beere HM. “The stress of dying”: The role of heat shock proteins in the regulation of apoptosis. J Cell Sci 2004; 117: 2641–2651.
[3] Nguyen L. Antibiotic resistance mechanisms in M. tuberculosis: an update. Arch Toxicol 2016; 90: 585–1604. [4] Hendrick JP, Hartl F. Functıons of Heat-Shock 1993.
[5] 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.
[6] Briffotaux J, Liu S, Gicquel B. Genome-wide transcriptional responses of Mycobacterium to antibiotics. Front Microbiol 2019; 10: 1–14.
[7] 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.
[8] 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.
[9] 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.
[10] 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.
[11] Guzel FD, Citak F. Development of an on-chip antibiotic permeability assay with single molecule detection capability. IEEE Trans Nanobioscience 2018; 17: 155–160.
[12] Malhotra S, Verma A, Tyagi N, Kumar V. Biosensors: principle, types and applications. Int J Adv Res Innov Ideas Educ 2017; 3: 3639–3644.
[13] Baselt DR, Lee GU, Natesan M, Metzger SW, Sheehan PE, Colton RJ. A biosensor based on magnetoresistance technology. Biosens Bioelectron 1998; 13: 731–739.
[14] Yang R, Jin J, Chen Y, Shao N, Kang H, Xiao Z, Tang Z, Wu Y, Zhu Z, Tan W. Carbon nanotube- quenched fluorescent oligonucleotides: probes that fluoresce upon hybridization. J Am Chem Soc 2008; 130: 8351–8358.
[15] Kaur J, Ghorbanpoor H, Öztürk Y, Kaygusuz Ö, Avcı H, Darcan C. Trabzon L, Güzel FD. On‐ chip label‐ free impedance‐ based detection of antibiotic permeation. IETNanobiotechnol 2021; 1-7.
[16] Niu SY, Li QY, Ren R, Zhang SS. Enzyme-enhanced fluorescence detection of DNA on etched optical fibers. Biosens Bioelectron 2009; 24: 2943–2946.
[17] Wilson JN, Teo YN, Kool ET. Efficient quenching of oligomeric fluorophores on a DNA backbone. J Am Chem Soc 2007; 129: 15426–15427.
[18] Randviir EP, Banks CE. Electrochemical impedance spectroscopy: an overview of bioanalytical applications. Anal Methods 2013; 5: 1098–1115.
[19] Blair EO, Corrigan DK. A review of microfabricated electrochemical biosensors for DNA detection. Biosens Bioelectron 2019; 134: 57–67.
[20] Liu J, Cao Z, Lu Y. Functional nucleic acid sensors. Chem Rev 2009; 109: 1948–1998.
[21] Blair EO, Hannah S, Vezza V, Avcı H, Kocagoz T, Hoskisson PA, Güzel FD, Corrigan DK. Biologically modified microelectrode sensors provide enhanced sensitivity for detection of nucleic acid sequences from Mycobacterium tuberculosis. Sensors and Actuators Reports. Sens. Actuator report 2020; 2: 100008.
[22] Huang Y, Bell MC, Suni II. Impedance biosensor for peanut protein Ara h 1. Anal Chem 2008; 80: 9157–9161.
[23] 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.
[24] 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.
[25] 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.
[26] PS J, Sutrave DS. A brief study of cyclic voltammetry and electrochemical analysis. Int J ChemTech Res 2018; 11: 77–88.
[27] Dizaji AN, Ali Z, Ghorbanpoor H, Ozturk Y, Akcakoca I, Avci H, Guzel FD. Electrochemical- based ‘‘antibiotsensor’’for the whole-cell detection of the vancomycin-susceptible bacteria. Talanta. 2021 Nov 1;234:122695.
[28] 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.
29] Rashid JIA, Yusof NA. The strategies of DNA immobilization and hybridization detection mechanism in the construction of electrochemical DNA sensor: A review. Sens Biosensing Res 2019; 16: 19-31.
[30] Bedford E. Gold surface nanostructuring for separation and sensing of biomolecules. Doctoral dissertation 2017.
[31] 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.
[32] Lisdat F, Schäfer D. The use of electrochemical impedance spectroscopy for biosensing. Anal Bioanal Chem 2008; 391: 1555–1567.
[33] Park JY, Park SM. DNA hybridization sensors based on electrochemical impedance spectroscopy as a detection tool. Sensors 2009; 9: 9513–9532.
[34] Drummond TG, Hill MG, Barton JK. Electrochemical DNA sensors. Nat Biotechnol 2003; l: 1192–1199.

APA	GUZEL F, AKCAKOCA I, GHORBANPOOR H, DIZAJI A, OZTURK Y, BLAIR E, KOCAGOZ T, CORRIGAN D (2021). HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. , 344 - 352. 10.18038/estubtda.806222
Chicago	GUZEL Fatma DOGAN,AKCAKOCA Iremnur,GHORBANPOOR Hamed,DIZAJI Araz Norouz,OZTURK Yasin,BLAIR Ewen O.,KOCAGOZ Tanil,CORRIGAN Damion K HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. (2021): 344 - 352. 10.18038/estubtda.806222
MLA	GUZEL Fatma DOGAN,AKCAKOCA Iremnur,GHORBANPOOR Hamed,DIZAJI Araz Norouz,OZTURK Yasin,BLAIR Ewen O.,KOCAGOZ Tanil,CORRIGAN Damion K HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. , 2021, ss.344 - 352. 10.18038/estubtda.806222
AMA	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. . 2021; 344 - 352. 10.18038/estubtda.806222
Vancouver	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. . 2021; 344 - 352. 10.18038/estubtda.806222
IEEE	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D "HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES." , ss.344 - 352, 2021. 10.18038/estubtda.806222
ISNAD	GUZEL, Fatma DOGAN vd. "HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES". (2021), 344-352. https://doi.org/10.18038/estubtda.806222

APA	GUZEL F, AKCAKOCA I, GHORBANPOOR H, DIZAJI A, OZTURK Y, BLAIR E, KOCAGOZ T, CORRIGAN D (2021). HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering, 22(4), 344 - 352. 10.18038/estubtda.806222
Chicago	GUZEL Fatma DOGAN,AKCAKOCA Iremnur,GHORBANPOOR Hamed,DIZAJI Araz Norouz,OZTURK Yasin,BLAIR Ewen O.,KOCAGOZ Tanil,CORRIGAN Damion K HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering 22, no.4 (2021): 344 - 352. 10.18038/estubtda.806222
MLA	GUZEL Fatma DOGAN,AKCAKOCA Iremnur,GHORBANPOOR Hamed,DIZAJI Araz Norouz,OZTURK Yasin,BLAIR Ewen O.,KOCAGOZ Tanil,CORRIGAN Damion K HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering, vol.22, no.4, 2021, ss.344 - 352. 10.18038/estubtda.806222
AMA	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering. 2021; 22(4): 344 - 352. 10.18038/estubtda.806222
Vancouver	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES. Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering. 2021; 22(4): 344 - 352. 10.18038/estubtda.806222
IEEE	GUZEL F,AKCAKOCA I,GHORBANPOOR H,DIZAJI A,OZTURK Y,BLAIR E,KOCAGOZ T,CORRIGAN D "HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES." Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering, 22, ss.344 - 352, 2021. 10.18038/estubtda.806222
ISNAD	GUZEL, Fatma DOGAN vd. "HIGHLY SENSITIVE LABEL-FREE ELECTROCHEMICAL DETECTION OF HEAT SHOCK PROTEIN WITH LOW-COST SCREEN-PRINTED ELECTRODES". Eskişehir Technical University Journal of Science and and Technology A- Applied Sciences and Engineering 22/4 (2021), 344-352. https://doi.org/10.18038/estubtda.806222