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通过表面等离子体共振对MUC16抗原/胺端基适配体相互作用进行超灵敏定量:动力学和热力学研究

Ultrasensitive Quantification of MUC16 Antigen/Amine-Terminated Aptamer Interaction by Surface Plasmon Resonance: Kinetic and Thermodynamic Studies.

作者信息

Valizadeh Shahbazlou Shahnam, Vandghanooni Somayeh, Dabirmanesh Bahareh, Eskandani Morteza, Hasannia Sadegh

机构信息

Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Adv Pharm Bull. 2024 Jul;14(2):388-399. doi: 10.34172/apb.2024.028. Epub 2024 Jan 13.

Abstract

PURPOSE

MUC16 is a commonly employed biomarker to identify and predict ovarian cancer (OC). Precise measurement of MUC16 levels is essential for the accurate diagnosis, prediction, and management of OC. This research seeks to introduce a new surface plasmon resonance (SPR) biosensor design that utilizes aptamer-based technology to enable the sensitive and real-time detection of MUC16.

METHODS

In this study, the sensor chip was immobilized with an anti-MUC16 aptamer (Ap) by utilizing 11-mercaptoundecanoic acid (MUA) as a linker to attach the amine-terminated Ap to the chip using EDC/NHS chemistry.

RESULTS

The results indicated that the newly created aptasensor had a detection limit of 0.03 U/mL for MUC16 concentration, with a linear range of 0.09 to 0.27 U/mL. The findings demonstrate good precision and accuracy (<15%) for each MUC16 concentration, with recoveries ranging from 93% to 96%. Additionally, the aptasensor exhibited high selectivity, good repeatability, stability, and applicability in real human serum samples, indicating its potential as a valuable tool for the diagnosis and treatment of OC.

CONCLUSION

According to the outcomes, the designed aptasensor exhibited acceptable specificity to detect the CA125 antigen and could be utilized for the serum detection of target antigen by SPR method.

摘要

目的

MUC16是一种常用于识别和预测卵巢癌(OC)的生物标志物。精确测量MUC16水平对于OC的准确诊断、预测和管理至关重要。本研究旨在引入一种新的表面等离子体共振(SPR)生物传感器设计,该设计利用基于适体的技术实现对MUC16的灵敏和实时检测。

方法

在本研究中,通过使用11-巯基十一烷酸(MUA)作为连接体,利用EDC/NHS化学方法将胺基末端的抗MUC16适体(Ap)固定在传感器芯片上。

结果

结果表明,新创建的适体传感器对MUC16浓度的检测限为0.03 U/mL,线性范围为0.09至0.27 U/mL。研究结果表明,对于每种MUC16浓度,该传感器都具有良好的精密度和准确度(<15%),回收率在93%至96%之间。此外,该适体传感器在实际人血清样本中表现出高选择性、良好的重复性、稳定性和适用性,表明其作为OC诊断和治疗的有价值工具的潜力。

结论

根据结果,所设计的适体传感器对检测CA125抗原具有可接受的特异性,可用于通过SPR方法对目标抗原进行血清检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3967/11347734/4ae5c8659fde/apb-14-388-g001.jpg

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