Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran.
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Biochim Biophys Acta Gen Subj. 2019 Oct;1863(10):1575-1582. doi: 10.1016/j.bbagen.2019.06.009. Epub 2019 Jun 19.
Here, we reported the development of a label-free and real-time surface plasmon resonance (SPR) based biosensor for cancer stem cells (CSCs) detection using cell surface biomarker; CD133. The fabricated biosensor was used for detection of this marker in some acute myeloid leukemia (AML) patients and the results were compared with those obtained from flow cytometry (FC) method. CD133 antibody was immobilized on the gold chip surface via EDC/NHS coupling method and binding of the candidate cells to the modified gold sensor surface was monitored after isolation of mononuclear cells from bone marrow of the patients. The method was validated in terms of various parameters such as CD133- antibody concentration and cell density. The CD133-marked cells were investigated in seven AML patients. All SPR results were compared with those obtained from FC method. A very good correlation (R = 0.96) was obtained between SPR and FC responses related to CD133-marked cells densities. In conclusion, in this study, a label-free and real-time SPR cytometry method was developed to detect CD133 and it was successfully applied to follow this cancer stem cell biomarker in AML patients.
在这里,我们报道了一种基于表面等离子体共振(SPR)的无标记和实时生物传感器的开发,用于使用细胞表面标志物 CD133 检测癌症干细胞(CSC)。该生物传感器用于检测一些急性髓系白血病(AML)患者中的这种标志物,并将结果与流式细胞术(FC)方法获得的结果进行比较。通过 EDC/NHS 偶联法将 CD133 抗体固定在金芯片表面,然后从患者骨髓中分离出单核细胞后,监测候选细胞与修饰后的金传感器表面的结合情况。该方法在 CD133-抗体浓度和细胞密度等各种参数方面进行了验证。在七名 AML 患者中研究了 CD133 标记的细胞。所有 SPR 结果均与 FC 方法获得的结果进行了比较。SPR 与 FC 反应之间与 CD133 标记细胞密度相关的相关性非常好(R=0.96)。总之,在这项研究中,开发了一种无标记和实时 SPR 细胞计数法来检测 CD133,并成功地应用于 AML 患者中这种癌症干细胞标志物的检测。