B-Type Natriuretic Peptide (BNP) Detection Using Electrochemical Immunosensor Based on Sandwich ELISA with Horseradish Peroxidase-Tetramethylbenzidine System

Procedia Technology Pub Date : 2017-01-01 DOI:10.1016/j.protcy.2017.04.065

Yeni Wahyuni Hartati , Ratna Nurmalasari , Shabarni Gaffar , Toto Subroto

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引用次数: 7

Abstract

A study of B-type natriuretic peptide (BNP) antigen as a good prognostic marker for patients with heart failure antigen detection employing an electrochemical immunosensor is described here. Monoclonal anti-BNP capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. An anti-BNP detection antibody conjugated with horseradish peroxidase enzyme (HRP) and 3,3,5,5’-tetramethybezidine dihydrochloride (TMB) was used as a substrate, in connection with voltammetric technique was achieve by measuring the peak current. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. Several optimizations were performed to reduce background signals; the optimal concentration of anti-BNP detection antibody needed, the time of SPCEs were first blocked with glycine, incubated with various concentrations of BNP and HRP-conjugated anti-BNP detection antibody (0 to 100.0 ng/mL), followed by electrochemical measurement were investigated,. In conclusion, this immunosensor greatly shortened and convenient for detection of heart failure diagnosis in real serum samples.

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基于夹心ELISA的电化学免疫传感器检测辣根过氧化物酶-四甲基联苯胺体系b型利钠肽

b型利钠肽(BNP)抗原作为心衰患者良好的预后标志物的研究，抗原检测采用电化学免疫传感器在这里描述。单克隆抗bnp捕获抗体固定在链霉亲和素修饰的spce上，以提高检测的灵敏度。以辣根过氧化物酶(HRP)和3,3,5,5′-四甲基bezidine dihydrochloride (TMB)偶联的抗bnp检测抗体为底物，结合伏安法测定峰值电流。链霉亲和素/生物素系统的结合导致捕获抗体定向良好的抗体固定化，从而提高了检测的灵敏度。进行了一些优化以减少背景信号;测定所需抗BNP检测抗体的最佳浓度，先用甘氨酸阻断spce，用不同浓度的BNP和酶标抗BNP检测抗体(0 ~ 100.0 ng/mL)孵育spce，然后进行电化学测定。综上所述，该免疫传感器大大缩短和方便了在真实血清样本中检测心力衰竭的诊断。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Procedia Technology

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