Materials Today Chemistry
Development of proof of concept for prostate cancer detection: an electrochemical immunosensor based on fullerene-C60 and copper nanoparticles composite film as diagnostic tool
Graphical abstract
Illustration of PSA sensing in serum sample via paper based immunosensor as a proof of concept.
Introduction
Providing diagnostic testing with high precision and sensitivity at the bedside of the patients is the need for clinical diagnostics and research as well. Therefore affordable, portable user-friendly, rapid and sensitive detection point of care (POC) testing devices are required to be developed [1]. Cancer is the leading cause of mortality and prostate cancer is a third major cause of death in men due to cancer [[2], [3], [4]]. However, currently, there is still no effective diagnostic tool for the early detection of prostate cancer. Early and accurate detection can provide the possibility to prolong survival time and increase the chances of successful treatment [[5], [6], [7]]. Prostate-specific antigen (PSA) is the most significant and effective protein biomarker for the screening of prostate cancer [8]. While diagnosing the patients for prostate cancer, total PSA concentration in human serum is determined. Increasing concentration of PSA beyond 4.0 ng/mL confirms the positive result of prostate cancer [9]. For the patients, who have been operated for prostate cancer, it is regarded as a serious sign when the level of PSA increases beyond 0.5 ng/mL [10]. In regard to these issues, sensitive and accurate detection of PSA becomes crucially necessary for early and quick detection and proper monitoring during treatment.
Nowadays, most of the preliminary diagnosis is being pursued and carried using extremely sophisticated techniques, such as computed and positron-emission tomography, ultrasound-based imaging, and nuclear magnetic resonance [11]. They entirely depend upon the growth and attainment of size of detectable cells. These methods are limited to the mass of billions of cells, but failure in the identification of advanced stage [12]. Apart from these, several advances in bioanalytical methods have been developed, such polymerase chain reaction (PCR) [13], enzyme-linked immunosorbent assay (ELISA) [14], mass spectroscopy (MS) [15], chromatography [16], electrophoresis [17], and surface plasmon-based methods [18]. These methods make the identification of a thousand times lesser-sized cells possible than previous conventional primary diagnosis. But the main disadvantages with these techniques are that they require sophisticated laboratory space, sample transportation, increased waiting times, increased administration and medical cost, required professional skilled technicians for the sample processing and handling conditions, and storage time, so sometimes it may produce inadequate results and finally increase the number of biopsies. Such problems may be overcome by using a simple and specific biosensing tool in the real-time analysis [19]. Though various types of biosensors have been developed, electrochemical immunosensors stand as a promising tool with several significant features such as high specificity, simplicity and biocompatibility, large surface area, less expensive with existing methods and easy to handle [20,21]. While developing immunosensing platforms, the stability of the immobilized proteins and tremendously increased signal intensity are the two key factors [22]. For efficient immobilization of bio proteins, better sensitivity, fast and facile detection of biomarkers various nanomaterials [8,[23], [24], [25], [26]] have been synthesized. Several metal nanoparticles are being used for the development of modified electrodes as an electrochemical sensor but copper nanoparticles (CuNPs) offer excellent properties due to small diameter, large specific surface area, and the ability for fast electron transfer. It is less expensive than other noble metals therefore, the application of copper nanoparticles in the development of modified electrodes is very promising [[27], [28], [29]].
Literature reveals that carbon-based materials have been extensively studied in a variety of applications [[30], [31], [32], [33], [34], [35]]. Various carbon-based materials such as carbon nanotubes (CNTs), graphene, and fullerene have drawn the attention of researchers for several important applications due to the high surface area, good electrical conductivity, strong mechanical strength, biocompatibility and low manufacturing cost. Recently, researchers have paid attention to fullerene-C60 due to its excellent electrochemical properties. The previous studies revealed that partially reduce fullerene-C60 films offer remarkably enhanced electrochemical behaviors in the aqueous solution therefore it has been used for modification electrodes for various electroactive molecules [[36], [37], [38]]. However, fullerene-C60 modified electrodes for the detection of analyte molecules are limited due to its sensitivity issue compared with other nanomaterials' modified electrodes. This could be overcome by incorporating metal nanoparticles over the fullerene-C60 film. However, metal nanoparticles-decorated fullerene-C60 modified electrodes for electrochemical applications are rarely reported in the literature [[39], [40], [41], [42]]. To the best of our knowledge this the first report on novel CuNP-reduced-fullerene-C60 composite film based immunosensor for PSA detection.
Present research work focuses upon the development of an accurate and simple immunosensing platform employing CuNP-reduced-fullerene-C60 nanocomposite film modified on the glassy carbon electrode (GCE). Excellent electron transfer kinetics and attachment of immobilized antibodies could increase at composite film due to enhanced biocompatibility and electrochemical properties. Due to a large amount of HRP-tagged secondary immobilized on CuNP-reduced-fullerene-C60 nanocomposite film, the electrochemical signal of fabricated immunosensor was greatly amplified achieving sensitive detection of PSA. Meanwhile proposed immunosensor also showed good reproducibility, long term stability, wide concentration range and lower detection limit. Therefore, the proposed immunosensor exhibited excellent potential for applications in the monitoring of prostate cancer.
Section snippets
Materials
Glassy carbon electrodes (GCE) were obtained from Sinsil International Pvt. Ltd. India. Anti-PSA-sc-7316 and HRP-tagged-sc-2060 were procured from Bio Square Biotechnologies India Pvt. Ltd. Prostate-specific antigen-PSA-sc-7316 was obtained from Sigma, India. 1-(3-(dimethylamino)-propyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxy succinimide (NHS), bovine serum albumin (BSA) and fullerene-C60, hydroquinone (HQ), potassium ferricyanide, potassium ferrocyanide, H2O2, Na2HPO4, NaH2PO4 from
Morphological characterization of CuNP-reduced-fullerene-C60/GCE electrode
The fabricated electrode was morphologically characterized by scanning electron microscopy (SEM). Fig. 1A–D represent SEM images of fullerene-C60, reduced fullerene-C60, CuNPs and CuNP-fullerene-C60 nanocomposite modified on ITO plates using the same method opted for GCE modification. Fig. 1B shows the scanning electron micrographs of fullerene-C60 after reduction in NaOH represents ultra-rod shape morphology with an association of bundle of micro rods. It can be noticed from Fig. 1D shows that
Conclusion
In summary, the present work describes the development of immunosensing platform based on fullerene-C60 and copper nanoparticles nanocomposite film for the electrochemical immunoassay of PSA. HQ@CuNP-reduced-fullerene-C60 nanocomposite film showed good biocompatibility, accelerated conductivity, the large surface area of sensing platform, which helped to immobilize Ab1 more efficiently. The high sensitivity of immunosensor could be attributed to the large surface area and excellent
Credit author statement
Lakkavarapu Suresh: Investigation process, acquisition and characterizations of the samples, data analysis, and drafting of the paper. Bondili Jayakumar Singh: Experiments on biological samples and supervision. Pradeep Kumar Brahman: Writing-review and editing, Conceptualization, validation and supervision
Declaration of competing interest
We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome.
Acknowledgments
The authors are thankful to Koneru Lakshmaiah Education Foundation, Vaddeswaram (A.P.) India, for providing necessary laboratory facilities. This work is financially supported by Science and Engineering Research Board, (Grant No. SB/FT/CS-019/2014) New Delhi, India, under its Fastrack Young Scientists Scheme and UGC, New Delhi under its UGC research award scheme for teachers (Grant no. 30-12/2015 SA-II).
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2023, BioelectrochemistryCitation Excerpt :In this regard, some substances with inherent redox activity have caught the eye of researchers. Suresh et al. [22] enriched copper nanoparticles (CuNPs) with fullerene -C60. Due to the synergistic effect of fullerene -C60 and CuNPs, it revealed remarkable catalytic activity for the reduction of hydrogen peroxide (H2O2), realizing rapid and trace detection of CEA.