Faculty of Chemistry, University of Warsaw, Pasteura Str. 1, PL 02-093 Warsaw, Poland; Faculty of Chemistry, Warsaw University of Technology, Noakowskiego Str. 3, PL 00-664 Warsaw, Poland.
Faculty of Chemistry, University of Warsaw, Pasteura Str. 1, PL 02-093 Warsaw, Poland.
Talanta. 2022 Sep 1;247:123600. doi: 10.1016/j.talanta.2022.123600. Epub 2022 May 28.
Monitoring the level of matrix metalloproteinase-9 (MMP-9) and inhibiting its expression is important for the diagnosis and treatment of various diseases. However, the analysis of MMP-9 is challenging owing to its very low content in the blood, especially at the early stages of diseases. Therefore, we developed an ultrasensitive and easy-to-use immunosensor based on a three-dimensional (3D) bioplatform for the determination of the total MMP-9 concentration in plasma. The used 3D bioplatform (G2 poly(amidoamine) dendrimer; PAMAM) improved the sensitivity of the determination by significantly expanding the surface area of the receptor layer. The antigen-antibody recognition process was controlled by quartz crystal microbalance with dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS). The effect of the orientation of antibody molecules in the sensing layer on the work parameters of the immunosensor was analyzed using unmodified PAMAM (PAMAM-NH) and PAMAM functionalized with -COOH groups (PAMAM-COOH). The developed immunosensor based on PAMAM-NH was characterized by a lower detection limit (LOD = 2.0 pg⋅mL) and wider analytical range (1·10 - 5 μg⋅mL for EIS and QCM-D) compared to PAMAM-COOH immunosensor (EIS: 1·10 - 0.5 μg⋅mL; QCM-D: 5·10 - 0.5 μg⋅mL). The functionality of the proposed device was verified in spiked plasma. The recoveries determined in commercial human and rat plasma and noncommercial rat plasma were very close to the value of 100% and in the range of 96-120% for Au/PAMAM-NH/Ab and Au/PAMAM-COOH/Ab immunosensors, respectively. The designed analytical devices showed high selectivity and sensitivity without the use of any amplifiers such as metal nanoparticles or enzymes.
监测基质金属蛋白酶-9（MMP-9）的水平并抑制其表达对于各种疾病的诊断和治疗非常重要。然而，由于其在血液中的含量非常低，尤其是在疾病的早期阶段，因此分析 MMP-9 具有挑战性。因此，我们开发了一种基于三维（3D）生物平台的超灵敏且易于使用的免疫传感器，用于测定血浆中总 MMP-9 浓度。所使用的 3D 生物平台（G2 聚（酰胺-胺）树枝状大分子；PAMAM）通过显著扩大受体层的表面积来提高测定的灵敏度。抗原-抗体识别过程通过石英晶体微天平（QCM-D）和电化学阻抗谱（EIS）进行控制。通过使用未修饰的 PAMAM（PAMAM-NH）和用-COOH 基团修饰的 PAMAM（PAMAM-COOH）分析了传感层中抗体分子取向对免疫传感器工作参数的影响。与 PAMAM-COOH 免疫传感器（EIS：1·10 - 0.5 μg·mL；QCM-D：5·10 - 0.5 μg·mL）相比，基于 PAMAM-NH 的开发的免疫传感器具有更低的检测限（LOD = 2.0 pg·mL -1 ）和更宽的分析范围（EIS 和 QCM-D 为 1·10 - 5 μg·mL -1 ）。在加标血浆中验证了所提出设备的功能。在商业人血浆和非商业大鼠血浆中测定的回收率非常接近 100%的值，并且对于 Au/PAMAM-NH/Ab 和 Au/PAMAM-COOH/Ab 免疫传感器分别在 96-120%的范围内。所设计的分析设备显示出高选择性和灵敏度，而无需使用任何放大器，例如金属纳米粒子或酶。
