Petre Shotadze Tbilisi Medical Academy, Tbilisi, Georgia.
Eur Rev Med Pharmacol Sci. 2024 Feb;28(4):1417-1422. doi: 10.26355/eurrev_202402_35463.
The Enzyme-Linked Immunosorbent Assay (ELISA) has been a cornerstone technique in laboratory medicine for over 55 years, relying on the specific binding of antibodies to antigens. ELISA's widespread use stems from its ability to detect low concentrations, its specificity, reproducibility, and potential for high-throughput screening. However, its sensitivity has limitations, prompting the exploration of innovative methods to improve the limit of detection (LOD). Nanoparticles provide a promising platform for enhancing ELISA sensitivity. Due to their high surface-to-volume ratio, they offer increased binding sites for capture elements and reporting tags, leading to amplified analytical signals. Recent studies have demonstrated improved sensitivity in ELISA through nanoparticle application, yielding faster detection times and enhanced sensitivities. This study investigates the potential of 50 nm citrate-capped silver nanoparticles to enhance ELISA's performance in quantifying cancer testis antigens (CTAs).
In our study, we used the Human NY-ESO-1 ELISA kit (for research purposes) to determine the concentration of CTAs in randomly selected samples from healthy (n=89) and oncological (n=80) subjects, aged 18-75. We employed 50 nm citrate-capped silver nanoparticles (AGCB50-1M, BioPure Silver Nanoparticles - bare citrate, nano-Composix, San Diego, CA, USA). ELISA reactions followed the manufacturer's instructions, and data processing aligned with the same guidelines. Absorbance (OD) measurements occurred at 450 nm, influencing nanoparticle selection. Each ELISA well contained 5 ml of nanoparticles' stock solution with specified concentrations. CTAs concentrations were derived from the standard curve through CurveExpert Basic software. Statistical analysis was performed using SPSS v. 27 software, with p-values indicating significance if <0.03. The study adhered to Helsinki Declaration principles and received ethical approval. Participants provided informed written consent.
The increased concentration values of CTAs for healthy individuals and cancer patients were determined in the case of the application of silver nanoparticles.
The usage of nanoparticles can enhance the sensitivity of the ELISA method and positively influence its specific detection limit.
酶联免疫吸附测定（ELISA）作为一种基于抗原抗体特异性结合的技术，已经在实验室医学领域中应用了超过 55 年。ELISA 的广泛应用源于其能够检测低浓度物质、具有特异性、重现性和高通量筛选的潜力。然而，其灵敏度存在一定的局限性，因此需要探索创新方法来提高检测限（LOD）。纳米颗粒为提高 ELISA 灵敏度提供了一个有前途的平台。由于其高的比表面积，它们为捕获元件和报告标签提供了更多的结合位点，从而产生放大的分析信号。最近的研究表明，通过纳米颗粒的应用可以提高 ELISA 的灵敏度，从而实现更快的检测时间和更高的灵敏度。本研究旨在探讨 50nm 柠檬酸包裹的银纳米颗粒在定量检测癌症睾丸抗原（CTA）方面对 ELISA 性能的增强作用。
在本研究中，我们使用了 Human NY-ESO-1 ELISA 试剂盒（用于研究目的）来确定随机选择的健康（n=89）和肿瘤（n=80）受试者（年龄 18-75 岁）血清中 CTA 的浓度。我们使用了 50nm 柠檬酸包裹的银纳米颗粒（AGCB50-1M，BioPure Silver Nanoparticles - bare citrate，nano-Composix，San Diego，CA，USA）。ELISA 反应按照制造商的说明进行，数据处理遵循相同的指南。在 450nm 处测量吸光度（OD），这影响了纳米颗粒的选择。每个 ELISA 孔中都含有 5ml 指定浓度的纳米颗粒储备液。通过 CurveExpert Basic 软件从标准曲线中得出 CTA 浓度。使用 SPSS v.27 软件进行统计分析，如果 p 值<0.03，则表示具有统计学意义。本研究遵循赫尔辛基宣言原则，并获得了伦理批准。参与者提供了书面知情同意。
在应用银纳米颗粒的情况下，确定了健康个体和癌症患者的 CTA 浓度值增加。
纳米颗粒的使用可以提高 ELISA 方法的灵敏度，并对其特定检测限产生积极影响。
