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1.
Adv Mater Interfaces ; 9(15): 2102526, 2022 May 23.
Article in English | MEDLINE | ID: mdl-35538925

ABSTRACT

The biofunctionalization of graphene field-effect transistors (GFETs) through vinylsulfonated-polyethyleneimine nanoscaffold is presented for enhanced biosensing of severe acute respiratory-related coronavirus 2 (SARS-CoV-2) spike protein and human ferritin, two targets of great importance for the rapid diagnostic and monitoring of individuals with COVID-19. The heterobifunctional nanoscaffold enables covalent immobilization of binding proteins and antifouling polymers while the whole architecture is attached to graphene by multivalent π-π interactions. First, to optimize the sensing platform, concanavalin A is employed for glycoprotein detection. Then, monoclonal antibodies specific against SARS-CoV-2 spike protein and human ferritin are anchored, yielding biosensors with limit of detections of 0.74 and 0.23 nm, and apparent affinity constants ( K D G F E T ) of 6.7 and 8.8 nm, respectively. Both biosensing platforms show good specificity, fast time response, and wide dynamic range (0.1-100 nm). Moreover, SARS-CoV-2 spike protein is also detected in spiked nasopharyngeal swab samples. To rigorously validate this biosensing technology, the GFET response is matched with surface plasmon resonance measurements, exhibiting linear correlations (from 2 to 100 ng cm-2) and good agreement in terms of K D values. Finally, the performance of the biosensors fabricated through the nanoscaffold strategy is compared with those obtained through the widely employed monopyrene approach, showing enhanced sensitivity.

2.
ACS Appl Mater Interfaces ; 13(36): 43696-43707, 2021 Sep 15.
Article in English | MEDLINE | ID: mdl-34470205

ABSTRACT

Graphene is a two-dimensional semiconducting material whose application for diagnostics has been a real game-changer in terms of sensitivity and response time, variables of paramount importance to stop the COVID-19 spreading. Nevertheless, strategies for the modification of docking recognition and antifouling elements to obtain covalent-like stability without the disruption of the graphene band structure are still needed. In this work, we conducted surface engineering of graphene through heterofunctional supramolecular-covalent scaffolds based on vinylsulfonated-polyamines (PA-VS). In these scaffolds, one side binds graphene through multivalent π-π interactions with pyrene groups, and the other side presents vinylsulfonated pending groups that can be used for covalent binding. The construction of PA-VS scaffolds was demonstrated by spectroscopic ellipsometry, Raman spectroscopy, and contact angle measurements. The covalent binding of -SH, -NH2, or -OH groups was confirmed, and it evidenced great chemical versatility. After field-effect studies, we found that the PA-VS-based scaffolds do not disrupt the semiconducting properties of graphene. Moreover, the scaffolds were covalently modified with poly(ethylene glycol) (PEG), which improved the resistance to nonspecific proteins by almost 7-fold compared to the widely used PEG-monopyrene approach. The attachment of recognition elements to PA-VS was optimized for concanavalin A (ConA), a model lectin with a high affinity to glycans. Lastly, the platform was implemented for the rapid, sensitive, and regenerable recognition of SARS-CoV-2 spike protein and human ferritin in lab-made samples. Those two are the target molecules of major importance for the rapid detection and monitoring of COVID-19-positive patients. For that purpose, monoclonal antibodies (mAbs) were bound to the scaffolds, resulting in a surface coverage of 436 ± 30 ng/cm2. KD affinity constants of 48.4 and 2.54 nM were obtained by surface plasmon resonance (SPR) spectroscopy for SARS-CoV-2 spike protein and human ferritin binding on these supramolecular scaffolds, respectively.


Subject(s)
Biomarkers/analysis , COVID-19/diagnosis , Graphite/chemistry , Immunoassay/methods , Spike Glycoprotein, Coronavirus/analysis , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Ethylenes/chemistry , Ferritins/immunology , Ferritins/metabolism , Humans , Point-of-Care Systems , Polyamines/chemistry , Polyethylene Glycols/chemistry , Pyrenes/chemistry , Quantum Theory , SARS-CoV-2/isolation & purification , SARS-CoV-2/metabolism , Semiconductors , Spike Glycoprotein, Coronavirus/immunology , Sulfonic Acids/chemistry , Surface Plasmon Resonance
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