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基于伏安法设计用于检测人绒毛膜促性腺激素的电化学生物传感器。

Designing an Electrochemical Biosensor Based on Voltammetry for Measurement of Human Chorionic Gonadotropin.

作者信息

Samare-Najaf Mohammad, Dehghanian Amirreza, Asadikaram Gholamreza, Mohamadi Maryam, Jafarinia Morteza, Savardashtaki Amir, Afshari Afrooz, Vakili Sina

机构信息

Blood Transfusion Research Center, Kerman Regional Blood Transfusion Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.

Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

J Med Signals Sens. 2024 Jul 25;14:21. doi: 10.4103/jmss.jmss_64_23. eCollection 2024.

Abstract

BACKGROUND

Human chorionic gonadotropin (hCG) is a polypeptide hormone synthesized during pregnancy and is also upregulated in some pathologic conditions such as certain tumors. Its measurement is essential for diagnosing pregnancy and malignancies. Despite numerous attempts to introduce an accurate method capable of detecting hCG levels, several limitations are found in previous techniques. This study aimed to address the limitations of current hCG assay methods by designing an electrochemical biosensor based on voltammetry for the rapid, selective, inexpensive, and sensitive measurement of hCG levels.

METHODS

A carbon paste electrode was prepared and functionalized by para-aminobenzoic acid. The primary anti-β-hCG monoclonal antibody was immobilized on the electrode surface by activating the carboxyl groups with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide solutions. The study also involved optimizing parameters such as the time for primary antibody fixation, the time for hCG attachment, and the pH of the hydrogen peroxide solution to maximize the biosensor response. Different concentrations of hCG hormone were prepared and loaded on the electrode surface, the secondary antibody labeled with HRP enzyme was applied, thionine in phosphate-buffered saline solution was placed on the electrode surface, and the differential pulse electrical signal was recorded.

RESULTS

The linear range ranged from 5 to 100 mIU/ml, and the limit of detection was calculated as 0.11 mIU. The relative standard deviation was 3% and 2% for five repeated measurements of commercial standard samples with concentrations of 2 and 20 mIU/mL, respectively. The percent recovery was obtained from 98.3% to 101.5%.

CONCLUSION

The sensor represents a promising advancement in hCG level measurement, offering a potential solution to overcome the existing limitations in current diagnostic strategies. Simple and inexpensive design, detecting hCG in its important clinical range during early pregnancy, and successful measurement of hCG in real serum samples are the advantages of this sensor.

摘要

背景

人绒毛膜促性腺激素(hCG)是一种在孕期合成的多肽激素,在某些病理状况如特定肿瘤中也会上调。其检测对于诊断妊娠和恶性肿瘤至关重要。尽管多次尝试引入能够检测hCG水平的准确方法,但先前技术存在若干局限性。本研究旨在通过设计一种基于伏安法的电化学生物传感器来解决当前hCG检测方法的局限性,以实现对hCG水平的快速、选择性、廉价且灵敏的测量。

方法

制备碳糊电极并用对氨基苯甲酸进行功能化。通过用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺和N-羟基琥珀酰亚胺溶液活化羧基,将抗β-hCG单克隆一抗固定在电极表面。该研究还涉及优化诸如一抗固定时间、hCG附着时间和过氧化氢溶液pH值等参数,以最大化生物传感器响应。制备不同浓度的hCG激素并加载到电极表面,应用辣根过氧化物酶(HRP)标记的二抗,将磷酸盐缓冲盐溶液中的硫堇置于电极表面,并记录差分脉冲电信号。

结果

线性范围为5至100 mIU/ml,检测限计算为0.11 mIU。对于浓度为2和20 mIU/mL的商业标准样品进行五次重复测量,相对标准偏差分别为3%和2%。回收率为98.3%至101.5%。

结论

该传感器在hCG水平测量方面代表了一项有前景的进展,为克服当前诊断策略中的现有局限性提供了潜在解决方案。设计简单且成本低廉、能够在妊娠早期检测hCG的重要临床范围以及成功测量真实血清样品中的hCG是该传感器的优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/abc1/11373787/362fc3783b6d/JMSS-14-21-g001.jpg

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