An origami paper-based electrochemical immunoassay for the C-reactive protein using a screen-printed carbon electrode modified with graphene and gold nanoparticles
An origami paper-based electrochemical immunoassay for C-reactive protein (CRP) detection is described. The assay integrates multiple steps of electrode modification into a single device. A graphene-modified screen-printed carbon electrode (G/SPCE) was employed to enhance sensitivity. Gold nanoparticles were first electrodeposited onto the G/SPCE, followed by a self-assembled monolayer of L-cysteine. The capture anti-CRP was then covalently immobilized on the modified electrode. CRP was quantified by measuring the changes in the charge-transfer resistance of the electrode by using hexacyanoferrate as the redox probe. Cyclic voltammetry and scanning electron microscopy were also applied to verify the successful modification of the electrode. Under optimal conditions, impedance increase in the 0.05–100 μg mL−1 CRP concentration range, and the limit of detection is 15 ng mL−1 (at S/N = 3). The immunoassay was successfully applied to the determination of CRP in a certified human serum sample. This method is simple, low-cost, portable and disposable.
KeywordsPaper-based analytical device Immunodetection Screen-printed carbon electrode Electrochemical impedance spectroscopy
This research was partially supported by The Center of Excellence on Petrochemical and Materials Technology (PETROMAT) through High Performance and Smart Materials (HPSM) research program. The authors also thank the Thailand Research Fund through Research Team Promotion Grant (RTA6080002), the Ratchadaphiseksomphot Endowment Fund under Outstanding Research Performance Program (SciSuperIII) and the Ratchadaphiseksomphot Endowment Fund of Chulalongkorn University for additional supports.
Compliance with ethical standards
The author(s) declare that they have no competing interests.
- 6.Kokkinos C, Prodromidis M, Economou A, Petrou P, Kakabakos S (2015) Disposable integrated bismuth citrate-modified screen-printed immunosensor for ultrasensitive quantum dot-based electrochemical assay of C-reactive protein in human serum. Anal Chim Acta 886:29–36. https://doi.org/10.1016/j.aca.2015.05.035 CrossRefPubMedGoogle Scholar
- 10.Charoenkitamorn K, Chaiyo S, Chailapakul O, Siangproh W (2018) Low-cost and disposable sensors for the simultaneous determination of coenzyme Q10 and α-lipoic acid using manganese (IV) oxide-modified screen-printed graphene electrodes. Anal Chim Acta 1004:22–31. https://doi.org/10.1016/j.aca.2017.12.026 CrossRefPubMedGoogle Scholar
- 11.Shen W, Tian D, Cui H, Yang D, Bian Z (2011) Nanoparticle-based electrochemiluminescence immunosensor with enhanced sensitivity for cardiac troponin I using N-(aminobutyl)-N-(ethylisoluminol)-functionalized gold nanoparticles as labels. Biosens Bioelectron 27(1):18–24. https://doi.org/10.1016/j.bios.2011.05.022 CrossRefPubMedGoogle Scholar
- 12.Byzova NA, Zherdev AV, Vengerov YY, Starovoitova ТA, Dzantiev BB (2017) A triple immunochromatographic test for simultaneous determination of cardiac troponin I, fatty acid binding protein, and C-reactive protein biomarkers. Microchim Acta 184(2):463–471. https://doi.org/10.1007/s00604-016-2022-1 CrossRefGoogle Scholar
- 14.Yan Q, Yang Y, Tan Z, Liu Q, Liu H, Wang P, Chen L, Zhang D, Li Y, Dong Y (2018) A label-free electrochemical immunosensor based on the novel signal amplification system of AuPdCu ternary nanoparticles functionalized polymer nanospheres. Biosens Bioelectron 103:151–157. https://doi.org/10.1016/j.bios.2017.12.040 CrossRefPubMedGoogle Scholar
- 15.Jampasa S, Siangproh W, Laocharoensuk R, Vilaivan T, Chailapakul O (2018) Electrochemical detection of c-reactive protein based on anthraquinone-labeled antibody using a screen-printed graphene electrode. Talanta 183:311–319. https://doi.org/10.1016/j.talanta.2018.02.075 CrossRefPubMedGoogle Scholar
- 18.Martinez AW, Phillips ST, Carrilho E, Thomas SW, Sindi H, Whitesides GM (2008) Simple telemedicine for developing regions: camera phones and paper-based microfluidic devices for real-time, off-site diagnosis. Anal Chem 80(10):3699–3707. https://doi.org/10.1021/ac800112r CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Pungjunun K, Chaiyo S, Jantrahong I, Nantaphol S, Siangproh W, Chailapakul O (2018) Anodic stripping voltammetric determination of total arsenic using a gold nanoparticle-modified boron-doped diamond electrode on a paper-based device. Microchim Acta 185(7):324. https://doi.org/10.1007/s00604-018-2821-7 CrossRefGoogle Scholar
- 25.Jampasa S, Siangproh W, Duangmal K, Chailapakul O (2016) Electrochemically reduced graphene oxide-modified screen-printed carbon electrodes for a simple and highly sensitive electrochemical detection of synthetic colorants in beverages. Talanta 160:113–124. https://doi.org/10.1016/j.talanta.2016.07.011 CrossRefPubMedGoogle Scholar
- 26.Wang L, Hua E, Liang M, Ma C, Liu Z, Sheng S, Liu M, Xie G, Feng W (2014) Graphene sheets, polyaniline and AuNPs based DNA sensor for electrochemical determination of BCR/ABL fusion gene with functional hairpin probe. Biosens Bioelectron 51:201–207. https://doi.org/10.1016/j.bios.2013.07.049 CrossRefPubMedGoogle Scholar
- 30.Feng D, Lu X, Dong X, Ling Y, Zhang Y (2013) Label-free electrochemical immunosensor for the carcinoembryonic antigen using a glassy carbon electrode modified with electrodeposited Prussian blue, a graphene and carbon nanotube assembly and an antibody immobilized on gold nanoparticles. Microchim Acta 180(9):767–774. https://doi.org/10.1007/s00604-013-0985-8 CrossRefGoogle Scholar