Electrochemical Immunosensor for Ultra-Low Detection of Human Papillomavirus Biomarker for Cervical Cancer

Abstract

Human papillomavirus (HPV) is the causative agent for cervical cancer. Of the various types of HPV, the high-risk HPV-16 type is the most important antigenic high-risk HPV. In this work, the antigenic HPV-16 L1 peptide was immobilized on a glassy carbon electrode and used to detect several concentrations of the anti-HPV-16 L1 antibody, and vice versa. Two electrode platforms were used: onion-like carbon (OLC) and its polyacrylonitrile (OLC-PAN) composites. Both platforms gave a wide linear concentration range (1.95 fg/mL to 6.25 ng/mL), excellent sensitivity (>5.2 μA/log ([HPV-16 L1, fg/mL]), and extra-ordinarily low limit of detection (LoD) of 1.83 fg/mL (32.7 aM) and 0.61 fg/mL (10.9 aM) for OLC-PAN and OLC-based immunosensors, respectively. OLC-PAN modified with the HPV-16 L1 protein showed low LoD for the HPV-16 L1 antibody (2.54 fg/mL, i.e., 45.36 aM), proving its potential use for screening purposes. The specificity of detection was proven with the anti-ovalbumin antibody (anti-OVA) and native ovalbumin protein (OVA). An immobilized antigenic HPV-16 L1 peptide showed insignificant interaction with anti-OVA in contrast with the excellent interaction with anti-HPV-16 L1 antibody, thus proving high specificity. The application of the immunosensor as a potential point-of-care (PoC) diagnostic device was investigated with screen-printed carbon electrodes, which detected ultra-low (ca. 0.7 fg/mL ≈ 12.5 aM) and high (ca. 12 μg/mL ≈ 0.21 μM) concentrations. This study represents the lowest LoD reported for HPV-16 L1. It opens the door for further investigation with other electrode platforms and realization of PoC diagnostic devices for screening and testing of HPV biomarkers for cervical cancer.

Ultrasensitive electrochemical immunosensor for the detection of C-reactive protein antigen

“…Subsequently, the physically absorbed antibodies were rinsed with PBS to remove the unbonded antibodies. The final coordination with the –NH 2 group of CRP-AB was obtained, with the amine-reactive sulfo-NHS ester groups on the working electrode, to form amide (–CONH–) bonds between the antibodies and the electrode, 44,45 which produced CRP-AB/OLC-PAN/GCE. Thereafter, any unreacted groups and nonspecific sites on the modified surface were deactivated and blocked by incubating the substrates in bovine serum albumin (BSA) 5% (w/v) for 2 h at 37 °C in a 0.1 M PBS (pH = 7.4), to prevent false positive binding.…”

Evaporation-Enhanced Redox Cycling for Rapid Detection of Attomolar SARS-CoV-2 Virions Using Nanolithography-free Electrochemical Devices

“…26,27 They can detect a wide range of viral targets, including the influenza virus, 28 hepatitis B virus, 29 and human papillomavirus. 30 As such, electrochemical sensors hold great promise as a powerful tool for the early diagnosis and management of viral infections. 31 Specifically, electrochemical devices can be miniaturized and integrated with readout electronics for simple-touse and high-throughput testing.…”

Ratiometric Electrochemiluminescence of Zirconium Metal–Organic Framework as a Single Luminophore for Sensitive Detection of HPV-16 DNA

“…Impressively, at different potentials, Zr-DPA@TCPP as a single luminophore displayed excellent dual ECL emissions, which were correspondingly derived from DPA and TCPP ligands. In addition, human papillomavirus (HPV) is a pathogen of diseases, and high-risk HPV-16 is considered to be responsible for cervical cancer cases . Therefore, the ratiometric ECL biosensor was effectively constructed and combined with DNA recycle amplification reactions to realize ultrasensitive detection of HPV-16 DNA (Scheme B).…”