Systematic Study of Various Functionalization Steps for Ultrasensitive Detection of SARS-CoV‑2 with Direct Laser-Functionalized Au-LIG Electrochemical Sensors

• Published in: ACS applied materials & interfaces 2024-09, Vol.16 (37), p.49041-49052
• Main Authors: Brustoloni, Caroline Ji-Mei, Soltan Khamsi, Pouya, Kammarchedu, Vinay, Ebrahimi, Aida
• Format: Article
• Language: English
• Subjects: antibodies; antigens; Betacoronavirus 1; Biological and Medical Applications of Materials and Interfaces; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; COVID-19 - diagnosis; COVID-19 - virology; COVID-19 infection; detection limit; diagnostic techniques; Electrochemical Techniques - instrumentation; Electrochemical Techniques - methods; electrochemistry; gold; Gold - chemistry; graphene; Graphite - chemistry; Humans; influenza; Lasers; Limit of Detection; Metal Nanoparticles - chemistry; nanomaterials; Nanostructures - chemistry; pandemic; SARS-CoV-2 - isolation & purification; Severe acute respiratory syndrome coronavirus 2; virion
• ISSN: 1944-8244; 1944-8252; 1944-8252
• DOI: 10.1021/acsami.4c09571

The 2019 coronavirus (COVID-19) pandemic impaired global health, disrupted society, and slowed the economy. Early detection of the infection using highly sensitive diagnostics is crucial in preventing the disease’s spread. In this paper, we demonstrate electrochemical sensors based on laser induced graphene (LIG) functionalized directly with gold (Au) nanostructures for the detection of SARS-CoV-2 with an outstanding limit of detection (LOD) of ∼1.2 ag·mL–1. To achieve the optimum performance, we explored various functionalization parameters to elucidate their impact on the LOD, sensitivity, and linearity. Specifically, we investigated the effect of (i) gold precursor concentration, (ii) cross-linker chemistry, (iii) cross-linker and antibody incubation conditions, and (iv) antigen–sensor interaction (diffusion-dominated incubation vs pipette-mixing), as there is a lack of a systematic study of these parameters. Our benchmarking analysis highlights the critical role of the antigen–sensor interaction and cross-linker chemistry. We showed that pipette-mixing enhances sensitivity and LOD by more than 1.6- and 5.5-fold, respectively, and also enables multimodal readout compared to diffusion-dominated incubation. Moreover, the PBA/Sulfo-NHS: EDC cross-linker improves the sensitivity and LOD compared to PBASE. The sensors demonstrate excellent selectivity against other viruses, including HCoV-229E, HCoV-OC43, HCoV-NL63, and influenza H5N1. Beyond the ability to detect antigen fragments, our sensors enable the detection of antigen-coated virion mimics (which are a better representative of the real infection) down to an ultralow concentration of ∼5 particles·mL–1.