Silane modified disposable ITO Electrode-based Biosensor for Sperm protein 17 detection as a Novel Cancer Biomarker

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Published Sep 24, 2021
Amit K. Yadav
Damini Verma Rinu Sharma Alok Thakkar Pratima R. Solanki

Abstract

Cancer has become a leading reason for deaths around the globe. The early detection using cancer-related biomarkers has provided critical knowledge on the progression of the disease, which is closely connected to successful cancer treatment [1]. Sperm protein 17 (SP17), an immunogenic mammalian protein found in the testis and spermatozoa, is a member of the cancer-testis antigen (CTA) family. SP17 has been identified as a tumor-associated antigen in ovarian cancer, multiple myeloma, head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer immunotherapy, and other human cancers [2-5]. The advent of selective, sensitive, and simple bio-sensing approaches for biomarker determination has piqued interest in clinical diagnosis [6], which is vital for assessing cancer status. The enzyme-linked immunosorbent assay, i.e., ELISA, has become one of the most dependable techniques today due to the specific, sensitive, reliable, and successful approach in hospitals and clinics all over the globe. Despite its many advantages, ELISA has some drawbacks, including a narrow dynamic range, high cost, excess reagent consumption, short stability of the labeled antibody, the requirement of centralized laboratory equipment, low sensitivity in the detection of biomolecular entities like microRNAs, and laborious assay/time-consuming procedure [7]. Furthermore, conventional ELISA's limit of detection (LOD) is close to the nanomolar concentration that becomes insufficient to fulfill the clinical threshold of numerous protein biomarkers, mainly in the initial stages of cancer [8]. Electrochemical biosensors are valuable instruments for clinical diagnostics and have recently gained a lot of attention because of their promising features like selectivity and sensitivity [9-11]. Due to the characteristics like compact in size, low-cost, and label-free, they have been commercialized for clinical, industrial, and environmental applications [12-15]. They are also appropriate for point-of-care (POC) applications. For the development of biosensors, monolayers of silane compounds are chemisorbed on commercially used indium tin oxide (ITO) or glass-modified electrodes. The epoxysilane molecule 3- glycidoxypropyltrimethoxysilane (GPTMS) is used to generate self-assembled monolayers on oxide surfaces without using the secondary crosslinker that occurred between the target antibody and silane molecule.

This study used a GPTMS modified ITO electrode to make a highly sensitive, selective, and label-free amperometric immunosensor for detecting SP17 in human serum. GPTMS was applied on the surface of the ITO electrode for anti-SP17 monoclonal antibody immobilization to design the immunosensor platform. This molecule contained an epoxy group that reacted with the amino groups of anti-SP17 antibodies, which were utilized as biomolecules for sensitive antigen contact. Using the cyclic voltammetry (CV) technique, a strong current signal was recorded in the presence of SP17 antigens. Field emission-scanning electron microscopy (FE-SEM), Atomic Force Microscopy (AFM), Raman, and Fourier transform infrared (FTIR) spectroscopy were used to undertake microscopic and spectroscopic surface characterizations of immunosensor fabrication processes. The immunosensor detected SP17 antigen with high sensitivity (0.047 mA pg mL-1 cm-2) and selectivity. In the detection range of 100-6000 pg mL-1, linearity was achieved by plotting a calibration curve between CV response and SP17 concentration, with the limit of detection (LOD) and limit of quantification (LOQ) of 47.57 pg mL-1 and 158.58 pg mL-1, respectively. Furthermore, because of the easy application methodology, it possessed strong repeatability, exceptional reproducibility, high stability, and reusability. It also displayed good recovery of SP17 antigen in real samples (human serum), indicating that the proposed immunosensor has a potential for SP17 detection in practical applications.

How to Cite

Yadav, A. K., Verma, D., Sharma, R., Thakkar, A., & Solanki, P. R. . (2021). Silane modified disposable ITO Electrode-based Biosensor for Sperm protein 17 detection as a Novel Cancer Biomarker. SPAST Abstracts, 1(01). Retrieved from https://spast.org/techrep/article/view/1164
Abstract 182 |

Article Details

Keywords

Immunosensor, Self-assembled monolayers, Cancer, Sperm protein 17, Biomarkers, Detection

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Section
SE1: Sensors

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