Electrochemical Biosensors Composed of Polyethylenimine (PEI) and Graphene Derivatives for Rapid Detection of Alzheimer’s Disease
Authors: Palley Bianca Fortes , de Arruda Milena Nakagawa , Souza Gustavo Freitas de , Artur Julio Cesar , Moraes Marli Leite de , Gonçalves Emerson Sarmento 
Published: February 2024

Graphene materials can be used as nanofillers in polymers to take advantage of graphene's suitable properties. Graphene-PEI composites are materials that combine the unique properties of graphene and PEI. These composites are made by combining graphene with PEI through physical or chemical method. The resulting material is characterized by enhanced mechanical properties, improved electrical conductivity and increased stability. By incorporating graphene into biosensor platforms, it can improve overall’s performance, including sensitivity, specificity and response. In this work, the detection of anti-βA40 biomarker was tested. Beta-amyloid peptide (βA1-40) was incorporated into liposomes of Dipalmitoyl Phosphatidylglycerol (DPPG) and was immobilized using Layer-by-Layer self-assembling technique on PEI_graphene derivatives composites. To obtain these composites it was utilized graphene oxide (GO) from Hummers’ method, it’s thermally reduced form (rGO) and also electrochemically exfoliated graphene (EEG). MilliQ water was added to the PEI to obtain a concentration of 2mg/mL and stirred for 30 min at room temperature for complete dissolution of the PEI. Then EEG, GO and rGO were added to PEI solution and subjected to ultrasonication in a Hielscher UP400ST sonicator, using the H14 probe for 30 minutes to obtain a homogeneous and stable dispersion.Carbon microelectrodes modified with PEI-graphene derivatives composites and (DPPG +Aβ-40) were used as biosensors platforms to detect anti-Aβ40 related to Alzheimer's disease. The assembly of the biosensors is shown in Figure 1. Cyclic Voltammetry was performed in three cycles, at a potential of -0.6 V to 0.6 V and scan rate of 0.05 V s-1 to evaluate the sensory response. The electrochemical measurements were performed with saline phosphate buffer solution (PBS). The morphology of PEI and PEI_graphene oxide can be seen in Figure 2. The large surface area of graphene allows for increased binding sites for bioreceptors, which may favor the adsorption of Aβ-40 peptide, improving signal transduction and sensitivity of the biosensor. The wettability of the composites was obtained through measurements of the static contact angle using the Ramé-Hart Contact Angle Goniometer (model 100-25-A). Figure 3 presents the contact angle for a film layer of the composites. All PEI_graphene derivative composites showed hydrophilic behavior (θ < 90°), PEI_EEG composite stood out as less hydrophilic (θ = 68°) among all composites. Hydrophilicity is essential in biosensors because it affects the interactions between the biological recognition elements and the target analytes, as well as the overall stability and reliability of the biosensor. Figure 4 presents the calibration curves using the normalized area obtained from cyclic voltammetry of the biosensors manufactured with 2 bilayers (PEI_graphene derivative/DPPG+Aβ40) and anti-Aβ40 biomarker in concentrations ranging from 1 ng mL to 5 µg mL-1. Among the tested PEI_graphene derivatives composites, the ones with EEG and RGO stood out in relation to pure PEI. Table 1 presents the adjusted R-square of the data in Figure 4. All composites, except PEI_GO, showed excellent linear fit, adjusted R-square close to 1 in the detection range between 1 and 1000 ng mL-1. The PEI_graphene oxide composite can be used in biosensors to enable rapid and efficient transduction of the binding signal, leading to improved detection performance.

Figure 1