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Sensitive detection of HSP70 using a current-amplified biosensor based on antibody-loaded PS-AuNPs@Cys/Au modified ITO chip

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Abstract

A biosensing electrochemical platform for heat shock protein 70 (HSP70) has been developed by integrating a three-electrode indium tin oxide (ITO) on a chip. The platform includes modifications to the reference electrode and working electrode for the detection of HSP70. The new platform is constructed by assembly of HSP70 antibody on PS-AuNPs@Cys/Au indium tin oxide (ITO) electrode to create a high HSP70 sensitive surface. The PS-AuNPs@Cys/Au indium tin oxide (ITO) electrode is obtained by immersing the ITO electrode into the PS-AuNPs@Cys solution and performing constant potential deposition at -1.4 V (Ag/AgCl). The PS-AuNPs@Cys/Au film deposited on ITO glass provides a desirable substrate for the immobilization of the HSP70 antibody and improves the loading of antibody between PS-AuNPs@Cys/Au and the electrode resulting in a significant amplification. Under optimal conditions, the fabricated sensor demonstrates a linear range extending from 0.1 ng mL− 1 to 1000 ng mL− 1, with an impressive detection limit of 25.7 pg mL− 1 (S/N = 3). The developed immunoassay method successfully detected the HSP70 content in normal human blood samples and outperformed the ELISA method commonly used for clinical sample analysis.

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References

  1. Guo Y, Shao S, Gu R, Hu X, Zhao M, Peng Y, Zhang W, Zhang B, Ding J, Wang N, Peng H, Han J (2023) Application of nanomaterials in early diagnosis of cancer. Nanotechnol Rev 12. https://doi.org/10.1515/ntrev-2023-0116

  2. Albakova Z, Mangasarova Y, Albakov A, Gorenkova L (2022) HSP70 and HSP90 in Cancer: cytosolic, endoplasmic reticulum and mitochondrial chaperones of Tumorigenesis. Front Oncol 12. https://doi.org/10.3389/fonc.2022.829520

  3. Gengyu Sha Z, Jiang W, Zhang C, Jiang D, Wang D, Tang (2023) The multifunction of HSP70 in cancer: Guardian or traitor to the survival of tumor cells and the next potential therapeutic target. Int Immunopharmacol. https://doi.org/10.1016/j.intimp.2023.110492

    Article  PubMed  Google Scholar 

  4. Rahal AK, Badgett RG, Hoffman RM (2016) Screening Coverage needed to reduce mortality from prostate Cancer: a living systematic review. PLoS ONE 11:e0153417. https://doi.org/10.1371/journal.pone.0153417

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Zhao K, Zhou G, Liu Y, Zhang J, Chen Y, Liu L, Zhang G (2023) HSP70 Family in Cancer: signaling mechanisms and therapeutic advances. Biomolecules 13:601. https://doi.org/10.3390/biom13040601

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Maher NG, Vergara IA, Long GV, Scolyer RA (2024) Prognostic and predictive biomarkers in melanoma. Pathol (Phila) 56:259–273. https://doi.org/10.1016/j.pathol.2023.11.004

    Article  CAS  Google Scholar 

  7. Chen J, Niu C, Yang N, Liu C, Zou S, Zhu S (2023) Biomarker discovery and application—An opportunity to resolve the challenge of liver cancer diagnosis and treatment. Pharmacol Res 189:106674. https://doi.org/10.1016/j.phrs.2023.106674

    Article  CAS  PubMed  Google Scholar 

  8. Guzhova IV, Margulis BA (2016) HSP70-based anti-cancer immunotherapy. Hum Vaccines Immunother. https://doi.org/10.1080/21645515.2016.1190057

    Article  Google Scholar 

  9. Zhang G, Liu Z, Ding H, Zhou Y, Doan HA, Sin KWT, Zhu ZJ, Flores R, Wen Y, Gong X, Liu Q, Li Y-P (2017) Tumor induces muscle wasting in mice through releasing extracellular Hsp70 and Hsp90. Nat Commun. https://doi.org/10.1038/s41467-017-00726-x

    Article  PubMed  PubMed Central  Google Scholar 

  10. Kasioumi P, Vrazeli P, Vezyraki P, Zerikiotis S, Katsouras C, Damalas A, Angelidis C (2018) Hsp70 (HSP70A1A) downregulation enhances the metastatic ability of cancer cells. Int J Oncol. https://doi.org/10.3892/ijo.2018.4666

    Article  PubMed  PubMed Central  Google Scholar 

  11. Lyu Q, Wawrzyniuk M, Rutten VPMG, van Eden W, Sijts AJAM, Broere F (2020) Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a canine macrophage cell line. Int J Mol Sci 21. https://doi.org/10.3390/ijms21186464

  12. Boudesco C, Cause S, Jego G, Garrido C (2018) Hsp70: a Cancer Target Inside and outside the cell, methods Mol. Biol Clifton NJ 1709:371–396. https://doi.org/10.1007/978-1-4939-7477-1_27

    Article  CAS  Google Scholar 

  13. Albakova Z, Armeev GA, Kanevskiy LM, Kovalenko EI, Sapozhnikov AM (2020) HSP70 Multi-functionality in Cancer. Cells 9. https://doi.org/10.3390/cells9030587

  14. Nycz M, Arkusz K, Pijanowska DG (2021) Fabrication of Electrochemical Biosensor based on Titanium Dioxide nanotubes and Silver nanoparticles for Heat shock protein 70 detection. Materials 14. https://doi.org/10.3390/ma14133767

  15. Maniya NH, Parashar K, Kadam LN, Srivastava DN (2021) Electrochemical detection of heat shock protein 70 over cost-effective plastic chip electrode platform. J Taiwan Inst Chem Eng 128:11–19. https://doi.org/10.1016/j.jtice.2021.09.009

    Article  CAS  Google Scholar 

  16. Feng F, Ataca ST, Ran M, Wang Y, Breen M, Kepler TB (2020) Gain-scanning for protein microarray assays. J Proteome Res 19:2664–2675. https://doi.org/10.1021/acs.jproteome.9b00892

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ayling K, Bowden T, Tighe P, Todd I, Dilnot EM, Negm OH, Fairclough L, Vedhara K (2017) The application of protein microarray assays in psychoneuroimmunology. Brain Behav Immun 59:62–66. https://doi.org/10.1016/j.bbi.2016.09.013

    Article  CAS  PubMed  Google Scholar 

  18. Aly NS, Kim H-S, Marei YM, Elhamshary AS, Bayoumi IR, Omar RE, Mohammed DA, Miyoshi S-I, Rashed GA (2023) Diagnosis of Toxoplasmosis using Surface Antigen Grade 1 detection by ELISA, Nano-Gold ELISA, and PCR in pregnant women. Int J Nanomed 18:1335–1345. https://doi.org/10.2147/IJN.S401876

    Article  CAS  Google Scholar 

  19. Sun B, Wang Y, Li D, Li W, Gou X, Gou Y, Hu F (2020) Development of a sensitive electrochemical immunosensor using polyaniline functionalized graphene quantum dots for detecting a depression marker. Mater Sci Eng C 111:110797. https://doi.org/10.1016/j.msec.2020.110797

    Article  CAS  Google Scholar 

  20. Sun B, Cai J, Li W, Gou X, Gou Y, Li D, Hu F (2018) A novel electrochemical immunosensor based on PG for early screening of depression markers-heat shock protein 70. Biosens Bioelectron 111:34–40. https://doi.org/10.1016/j.bios.2018.03.049

    Article  CAS  PubMed  Google Scholar 

  21. Sonuç Karaboğa MN, Şimşek ÇS, Sezgintürk MK (2016) AuNPs modified, disposable, ITO based biosensor: early diagnosis of heat shock protein 70. Biosens Bioelectron 84:22–29. https://doi.org/10.1016/j.bios.2015.08.044

    Article  CAS  PubMed  Google Scholar 

  22. Özcan B, Sezgintürk MK (2016) Graphene oxide based electrochemical label free immunosensor for rapid and highly sensitive determination of tumor marker HSP70. Talanta 160:367–374. https://doi.org/10.1016/j.talanta.2016.07.039

    Article  CAS  PubMed  Google Scholar 

  23. Can F, Akkas T, Bekler SY, Takmakli S, Uzun L, Ozaydin G, Ince (2024) Selective determination of an ovarian cancer biomarker at low concentrations with surface imprinted nanotube based Chemosensor. Bioelectrochemistry 157:108655. https://doi.org/10.1016/j.bioelechem.2024.108655

    Article  CAS  PubMed  Google Scholar 

  24. Ma S, Zhao W, Liu X, Li Y, Ma P, Zhang K, Zhang Q (2024) A novel microfluidic chip integrating with microcolumn array electrodes for rapid and ultrasensitive detection of alpha-fetoprotein. Anal Chim Acta 1291:342240. https://doi.org/10.1016/j.aca.2024.342240

    Article  CAS  PubMed  Google Scholar 

  25. Ionescu R, Selon R, Pocholle N, Zhou L, Rumyantseva A, Bourillot E, Lesniewska E (2018) Microwave Spectroscopic Detection of Human Hsp70 protein on annealed gold nanostructures on ITO Glass strips. Biosensors 8:118. https://doi.org/10.3390/bios8040118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Chen D, Song Z, Lian M, Yang Y, Lin S, Xiao L (2021) Single-particle fibrinogen detection using platelet membrane-coated fluorescent polystyrene nanoparticles. Nanoscale 13:2914–2922. https://doi.org/10.1039/D0NR08492A

    Article  CAS  PubMed  Google Scholar 

  27. Sun J, Wang R, Wang L, Wang X, Wang J, Shi Z, Chen Z, Wang M, Xu C (2023) Visual/quantitative SERS biosensing chip based on Au-decorated polystyrene sphere microcavity arrays. Sens Actuators B Chem 388:133869. https://doi.org/10.1016/j.snb.2023.133869

    Article  CAS  Google Scholar 

  28. Chen B, Yu Q, Chen Z, Zhu W, Li S, You H, Lv Z, Liu Y, Hu Q, Zheng Z, Farhana Y (2021) Polystyrene microsphere assisted synthesis of a Co/PEG comodified PbO2 anode and its electrocatalytic oxidation performance. Sep Purif Technol 279:119792. https://doi.org/10.1016/j.seppur.2021.119792

    Article  CAS  Google Scholar 

  29. Choi S, Vazquez-Duhalt R, Graeve OA (2022) Nonlinear charge regulation for the deposition of silica nanoparticles on polystyrene spherical surfaces. J Colloid Interface Sci 613:747–763. https://doi.org/10.1016/j.jcis.2022.01.076

    Article  CAS  PubMed  Google Scholar 

  30. Wang H, Ma R, Nienhaus K, Nienhaus GU (2019) Formation of a monolayer protein Corona around Polystyrene nanoparticles and implications for Nanoparticle Agglomeration. SMALL 15. https://doi.org/10.1002/smll.201900974

  31. Liao Y, An M, Hao X, Song X, Yang Z, Ren H, Liu Z (2021) Enhanced floatability of low rank coal using surface functionalized polystyrene nanoparticles as collectors. J Clean Prod 284:124763. https://doi.org/10.1016/j.jclepro.2020.124763

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to appreciate the financial support, in whole or in part, from the National Natural Science Foundation of China (Nos. 61071026, 31771079).

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Authors and Affiliations

  1. School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, 610054, PR China

    Ruming Liu, Yan Liu, Chaoyu Li, Dorothy Araba Yakoba Agyapong, Juan Feng, Lixia Tang & Hongjuan Zeng

  2. Biomedical Engineering Program, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana

    Dorothy Araba Yakoba Agyapong

Authors
  1. Ruming Liu
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  2. Yan Liu
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  3. Chaoyu Li
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  4. Dorothy Araba Yakoba Agyapong
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  5. Juan Feng
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  7. Hongjuan Zeng
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Contributions

Ruming Liu: Data curation, Formal analysis, Visualization, Writing-original draft, review & editing. Yan Liu: Writing-review & editing. Chaoyu Li: Writing-review & editing. Dorothy Araba Yakoba Agyapong: Validation, Writing-review & editing. JuanFeng: Validation Lixia Tang: Validation, Writing-review & editing. Hongjuan Zeng: Conceptualization, Supervision, Funding acquisition.

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Correspondence to Hongjuan Zeng.

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Liu, R., Liu, Y., Li, C. et al. Sensitive detection of HSP70 using a current-amplified biosensor based on antibody-loaded PS-AuNPs@Cys/Au modified ITO chip. Microchim Acta 191, 272 (2024). https://doi.org/10.1007/s00604-024-06333-0

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