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适配体-抗体双探针在单壁碳纳米管桥接介电电极上的应用:对人凝血因子的比较分析。

Aptamer-antibody dual probes on single-walled carbon nanotube bridged dielectrode: Comparative analysis on human blood clotting factor.

机构信息

Emergency Department, The Affiliated Hospital of Qingdao University, Qingdao 266000, PR China.

Department of Radiology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province 250013, PR China.

出版信息

Int J Biol Macromol. 2020 May 15;151:1133-1138. doi: 10.1016/j.ijbiomac.2019.10.156. Epub 2019 Nov 16.

DOI:10.1016/j.ijbiomac.2019.10.156
PMID:31743722
Abstract

Haemophilia is a blood clotting disorder known as 'Christmas disease' caused when the blood has defect with the clotting factor(s). Bleeding leads various issues, such as chronic pain, arthritis and a serious complication during the surgery. Identifying this disease is mandatory to take the necessary treatment and maintains the normal clotting. It has been proved that the level of factor IX (FIX) is lesser with haemophilia patient and the attempt here is focused to quantify FIX level by interdigitated electrode (IDE) sensor. Single-walled carbon nanotube (SWCNT) was utilized to modify IDE sensing surface. On this surface, dual probing was evaluated with aptamer and antibody to bring the possible advantages. The detection limit with antibody was found to be 1 pM, while aptamer shows 100 fM. Further, a fine-tuning was attempted with sandwich pattern of aptamer-FIX-antibody and antibody-FIX-aptamer and compared. Specific elevation of detection with 10 folds was noticed and displayed the detection at 100 f. in both sandwich patterns. In addition, FIX was detected in the diluted human serum by aptamer-FIX-antibody sandwich, it was found that FIX detected from the dilution factor 1:640. A novel demonstration is with higher discrimination against other clotting factors, XI and VII.

摘要

血友病是一种凝血障碍疾病,被称为“圣诞节病”,是由于血液中凝血因子(s)存在缺陷引起的。出血会导致各种问题,如慢性疼痛、关节炎以及手术中的严重并发症。因此,必须明确诊断这种疾病,以便采取必要的治疗措施并维持正常的凝血功能。已经证明血友病患者的因子 IX(FIX)水平较低,本研究旨在通过叉指电极(IDE)传感器来定量 FIX 水平。单壁碳纳米管(SWCNT)被用于修饰 IDE 传感表面。在该表面上,评估了适配子和抗体的双重探测,以带来可能的优势。发现抗体的检测限为 1 pM,而适配子的检测限为 100 fM。此外,还尝试了适配子-FIX-抗体和抗体-FIX-适配子的三明治模式的微调,并进行了比较。在两种三明治模式中,都观察到了 10 倍的检测灵敏度提高,并显示出 100 fM 的检测限。此外,通过适配子-FIX-抗体三明治法在稀释的人血清中检测到了 FIX,发现从稀释因子 1:640 可以检测到 FIX。此外,本研究还对其他凝血因子 XI 和 VII 具有更高的区分能力。

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