Institute of Ecology and Genetics of Microorganisms, Urals Branch of RAS, Perm, Russia.
Biology Faculty, Perm State University, Perm, Russia.
Analyst. 2024 Nov 18;149(23):5657-5667. doi: 10.1039/d4an01160k.
LaNiO perovskite nanoparticles, especially nanospheres (LNNS), show great promise in biomedical assays due to their peroxidase-like catalytic properties. However, LNNS-based diagnostic reagents have not been tested in nanozyme enzyme-linked immunosorbent assay (NLISA) or other enzyme-linked immunosorbent assays, and there is limited data on their synthesis. To fill this gap, it is necessary to develop a method for creating LNNS conjugates with monoclonal antibodies and to investigate the reproducibility, scalability, and applicability of these diagnostic reagents in NLISA. We have successfully developed a method for producing novel diagnostic reagents utilizing LaNiO nanospheres. Our research demonstrates the application of these nanospheres in a NLISA specifically designed for the detection of C-reactive protein (CRP) in real serum samples. This method is both reproducible and scalable, allowing for the efficient production of nanospheres that are functionalized with monoclonal antibodies targeting CRP, with a mean diameter of approximately 270 nm. Based on the promising results obtained from our experiments, we have developed and optimized a sandwich-format NLISA for CRP detection. This assay achieved a lower limit of detection at 0.178 μg L, with a dynamic range from 12.5 to 0.195 μg L and a linear detection range extending from 0.195 to 6.25 μg L, showcasing its potential for clinical applications. The new NLISA method, utilizing LaNiO nanospheres in a sandwich format for the detection of CRP, significantly enhances sensitivity compared to similar use horseradish peroxidase-based ELISA. In this study for the first time, the functionalization of lanthanum nickelate nanospheres with recognition elements has been demonstrated. This advancement also sheds light on the technological challenges involved in synthesizing diagnostic reagents, identifying areas that need further exploration.
镧镍矿纳米粒子，特别是纳米球（LNNS），由于其过氧化物酶样催化特性，在生物医学分析中具有很大的应用前景。然而，基于 LNNS 的诊断试剂尚未在纳米酶酶联免疫吸附测定（NLISA）或其他酶联免疫吸附测定中进行测试，并且关于其合成的信息有限。为了填补这一空白，有必要开发一种将 LNNS 与单克隆抗体偶联的方法，并研究这些诊断试剂在 NLISA 中的重现性、可扩展性和适用性。我们已经成功开发了一种利用 LaNiO 纳米球制备新型诊断试剂的方法。我们的研究表明，这些纳米球在专门设计用于检测实际血清样本中 C-反应蛋白（CRP）的 NLISA 中得到了应用。该方法具有重现性和可扩展性，允许高效生产功能化的纳米球，这些纳米球通过 CRP 的单克隆抗体进行功能化，平均直径约为 270nm。基于我们实验中获得的有前景的结果，我们已经开发并优化了用于 CRP 检测的夹心型 NLISA。该测定法的检测下限为 0.178μg/L，动态范围为 12.5-0.195μg/L，线性检测范围从 0.195-6.25μg/L 扩展，展示了其在临床应用中的潜力。新的 NLISA 方法，在夹心型中利用 LaNiO 纳米球检测 CRP，与基于辣根过氧化物酶的 ELISA 相比，显著提高了灵敏度。在这项研究中，首次证明了镧镍酸盐纳米球的功能化与识别元件的结合。这一进展还揭示了在合成诊断试剂方面所涉及的技术挑战，确定了需要进一步探索的领域。
