RESUMO
Helicobacter pylori (H. pylori) infection is highly prevalent worldwide, affecting more than 43% of world population. The infection can be transmitted through different routes, like oral-oral, fecal-oral, and gastric-oral. Electrochemical sensors play a crucial role in the early detection of various substances, including biomolecules. In this study, the development of nanobody (Nb)-based immunosensor for the detection of H. pylori antigens in saliva samples was investigated. The D2_Nb was isolated and characterized using Western blot and ELISA and employed in the fabrication of the immunosensor. The sensor was prepared using gold screen-printed electrodes, with the immobilization of Nb achieved through chemical linkage using cysteamine-glutaraldehyde. The surface of the electrode was characterized using EIS, FTIR and SEM. Initially, the Nb-based immunosensor's performance was evaluated through cyclic voltammetry (CV), differential pulse voltammetry (DPV), and square wave voltammetry (SWV). The sensor exhibited excellent linearity with an R2 value of 0.96. However, further assessment with the DPV technique revealed both a low limit of detection (5.9 ng/mL, <1 cfu/mL) and high selectivity when exposed to a mixture of similar antigens. Moreover, the immunosensor demonstrated robust recovery rates (96.2%-103.4%) when spiked into artificial saliva and maintained its functionality when stored at room temperature for 24 days.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Infecções por Helicobacter , Helicobacter pylori , Limite de Detecção , Saliva , Anticorpos de Domínio Único , Saliva/microbiologia , Saliva/química , Técnicas Biossensoriais/instrumentação , Helicobacter pylori/imunologia , Helicobacter pylori/isolamento & purificação , Humanos , Anticorpos de Domínio Único/química , Anticorpos de Domínio Único/imunologia , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/imunologia , Imunoensaio/métodos , Ouro/química , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/análise , Antígenos de Bactérias/isolamento & purificaçãoRESUMO
In this study, a hybrid material, MIL-53(Al)@CBS, was synthesized via the solvothermal method, involving the growth of MIL-53(Al) crystals on cocoa bean shell residues (CBS). Physicochemical characterization techniques, including TGA, BET, FTIR, XRD, and SEM, confirmed successful hybridization. MIL-53(Al)@CBS was employed as an adsorbent for antibiotics (oxytetracycline, tetracycline, chlortetracycline) separation from aqueous solutions. Parameters like pH, adsorbent dose, concentration, time, and temperature were systematically evaluated. FTIR revealed π-π interactions and hydrogen bonds between tetracyclines and the adsorbent. MIL-53(Al)@CBS exhibited adsorption, with removal rates up to 98.92%, 99.04%, and 98.24% for OTC, TC, and CTC, respectively. Kinetics suggested adsorption depends on active site availability, with TC adsorbing fastest. Microscopic models showed adsorption on three distinct active site types with different affinities without competition or adherence to the Langmuir hypothesis. Importantly, MIL-53(Al)@CBS maintained high adsorption capacity even after ten washing cycles, highlighting its potential for water treatment.