口腔癌是主要的癌症类型之一，通常在晚期才被诊断出来，导致患者预后不良，治疗选择也较少。为了解决这一差距，利用抗生物污染水凝胶的生物传感器在非侵入性体液中进行早期口腔癌检测变得越来越重要。在此，我们展示了一种创新的电化学免疫传感器的制造，用于快速、无标记、非侵入性且经济实惠地检测肿瘤坏死因子-α (TNF-α)，这是人工唾液样本中与口腔癌进展相关的生物标志物。金丝网印刷电极（gSPE）采用绿色合成的多孔电活性还原氧化石墨烯（rGO）水凝胶进行修饰，利用L-胱氨酸（L-cys）作为原位还原剂和表面功能化剂，然后共价固定抗-TNF-α并用牛血清白蛋白（BSA）封闭残留位点以制造BSA/抗TNF-α/L-cys_rGO水凝胶/gSPE免疫传感平台。所制造的平台表现出优异的性能，检测下限为 1.20 pg mL-1，线性范围为 1 至 200 pg mL-1，灵敏度高达 2.10 μA pg-1 mL cm-2。差分脉冲伏安法（DPV）技术。此外，即使存在潜在干扰物和其他癌症生物标志物，它也对 TNF-α 表现出特异性。此外，生物传感器表现出良好的重现性和重复性，相对标准偏差（%RSD）分别为5.11%和1.85%。因此，将 L-cys_rGO 水凝胶集成到免疫传感器设计中可提供增强的性能，为其在早期口腔癌诊断中的应用铺平道路。

Oral cancer is one of the leading cancer types, which is frequently diagnosed at an advanced stage, giving patients a poor prognosis and fewer therapeutic choices. To address this gap, exploiting biosensors utilizing anti-biofouling hydrogels for early-stage oral cancer detection in non-invasive body fluids is gaining utter importance. Herein, we have demonstrated the fabrication of an innovative electrochemical immunosensor for the rapid, label-free, non-invasive, and affordable detection of tumor necrosis factor-α (TNF-α), a biomarker associated with oral cancer progression in artificial saliva samples. The gold screen-printed electrodes (gSPEs) are modified with a green synthesized porous and electroactive reduced graphene oxide (rGO) hydrogel utilizing L-cystine (L-cys) as both in situ reducing and surface functionalization agent, followed by covalent immobilization of anti-TNF-α and blocking of residual sites with bovine serum albumin (BSA) to fabricate the BSA/anti-TNF-α/L-cys_rGO hydrogel/gSPE immunosensing platform. The fabricated platform demonstrates excellent performance, with a low limit of detection of 1.20 pg mL-1, a broad linear range from 1 to 200 pg mL-1, and a high sensitivity of 2.10 μA pg-1 mL cm-2 carried out with differential pulse voltammetry (DPV) technique. Moreover, it exhibits specificity towards TNF-α, even in the presence of potential interferents and other cancer biomarkers. Besides, the biosensor showed good reproducibility and repeatability with a relative standard deviation (%RSD) of 5.11% and 1.85%, respectively. Thus, integrating the L-cys_rGO hydrogel in the immunosensor design offers enhanced performance, paving the way for its application in early-stage oral cancer diagnosis.