Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-154, Iran.
Analytical Chemistry Research Laboratory, Mobin Shimi Azma Company, Tehran, Iran.
J Nanobiotechnology. 2020 Jun 17;18(1):93. doi: 10.1186/s12951-020-00648-9.
Since vascular endothelial growth factor (VEGF) is a significant regulator of cancer angiogenesis, it is essential to develop a technology for its sensitive detection. Herein, we sensitized a chemiluminescence (CL) immunoassay through the combination of HO-sensitive TGA-CdTe quantum dot (QD) as signal transduction, dextran as a cross-linker to prepare enzyme-labeled antigen and the ultrahigh bioactivity of catalase (CAT) as reporter enzyme.
Under the optimized experimental conditions, the chemiluminescence enzyme-linked immunosorbent assay (CL-ELISA) method can detect VEGF in the excellent linear range of 2-35,000 pg mL, with a detection limit (S/N = 3) of 0.5 pg mL which was approximately ten times lower than the commercial colorimetric immunoassay. This proposed method has been successfully applied to the clinical determination of VEGF in the human serum samples, and the results illustrated an excellent correlation with the conventional ELISA method (R = 0.997). The suitable recovery rate of the method in the serum ranged from 97 to 107%, with a relative standard deviation of 1.2% to 13.4%.
The novel immunoassay proposes a highly sensitive, specific, and stable method for very low levels detection of VEGF that can be used in the primary diagnosis of tumors. With the well-designed sensing platform, this approach has a broad potential to be applied for quantitative analysis of numerous disease-related protein biomarkers for which antibodies are available.
由于血管内皮生长因子 (VEGF) 是癌症血管生成的重要调节剂,因此开发一种用于其敏感检测的技术至关重要。在此,我们通过将 HO 敏感的 TGA-CdTe 量子点 (QD) 用作信号转导、葡聚糖作为交联剂来制备酶标记抗原,以及超高效生物活性的过氧化氢酶 (CAT) 作为报告酶,使化学发光 (CL) 免疫分析变得敏感。
在优化的实验条件下,化学发光酶联免疫吸附测定 (CL-ELISA) 方法可以在 2-35,000 pg mL 的优异线性范围内检测 VEGF,检测限 (S/N = 3) 为 0.5 pg mL,大约比商业比色免疫测定低十倍。该方法已成功应用于人血清样品中 VEGF 的临床测定,结果与常规 ELISA 方法具有极好的相关性 (R = 0.997)。该方法在血清中的回收率适宜范围为 97%至 107%,相对标准偏差为 1.2%至 13.4%。
该新型免疫分析为检测 VEGF 提供了一种高灵敏度、特异性和稳定性的方法,可用于肿瘤的初步诊断。该方法具有广阔的应用潜力,可用于定量分析许多具有抗体的疾病相关蛋白质生物标志物。