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氧化锌薄膜中的抗体固定化作为一种易于操作的检测策略

Antibody Immobilization in Zinc Oxide Thin Films as an Easy-Handle Strategy for Detection.

作者信息

Salinas Domínguez Rafael Antonio, Domínguez Jiménez Miguel Ángel, Orduña Díaz Abdú

机构信息

Centro de Investigación en Biotecnología Aplicada del Instituto Politécnico Nacional (CIBA-IPN), Tlaxcala 72197, Mexico.

Centro de Investigaciones en Dispositivos Semiconductores, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla (BUAP), Puebla 72570, Mexico.

出版信息

ACS Omega. 2020 Aug 10;5(32):20473-20480. doi: 10.1021/acsomega.0c02583. eCollection 2020 Aug 18.

DOI:10.1021/acsomega.0c02583
PMID:32832800
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7439397/
Abstract

The antibody immobilization compatible with low-cost materials and label-free strategies is a challenge for biosensor device fabrication. In this study, ZnO thin film deposition was carried out on corning glass substrates by ultrasonic spray pyrolysis at 200 °C. The thin films were analyzed as platforms for enteropathogenic ( EPEC) antibody immobilization. The modification of thin films from the functionalization and antibody immobilization steps was visualized using Fourier transform infrared spectroscopy (FTIR) spectroscopy, and surface changes were observed by atomic force microscopy. The obtained FTIR spectra after functionalization showed a contribution of the amino group (NH) derived from silane (3-aminopropyltrimethoxysilane). The antibody immobilization showed an amide I conserved signal corresponding to the C=O stretching vibrations and the amide II signal related to the N-H scissor vibration mode. In this way, the signals observed are correlated with the presence of antibody immobilized on the film. The ZnO film morphology changes after every stage of the process and allows observing the antibody distribution on the immobilized surface. In order to validate the antibody recognition capability as well as the EPEC detection , polymerase chain reaction was used.

摘要

与低成本材料和无标记策略兼容的抗体固定化是生物传感器装置制造面临的一项挑战。在本研究中，通过超声喷雾热解法于200℃在康宁玻璃基板上进行了ZnO薄膜沉积。这些薄膜被分析作为肠道致病性大肠杆菌（EPEC）抗体固定化的平台。使用傅里叶变换红外光谱（FTIR）对功能化和抗体固定化步骤中薄膜的改性进行可视化分析，并通过原子力显微镜观察表面变化。功能化后获得的FTIR光谱显示出源自硅烷（3-氨丙基三甲氧基硅烷）的氨基（NH）的贡献。抗体固定化显示出与C=O伸缩振动相对应的酰胺I保守信号以及与N-H剪式振动模式相关的酰胺II信号。通过这种方式，观察到的信号与固定在薄膜上的抗体的存在相关。该过程每个阶段后ZnO薄膜的形态都会发生变化，并且能够观察到抗体在固定表面上的分布。为了验证抗体识别能力以及EPEC检测，使用了聚合酶链反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/b2946dadb87b/ao0c02583_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/7a24d88ea4d2/ao0c02583_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/35a6753254e7/ao0c02583_0002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/6c9db402c76a/ao0c02583_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/cbbe2838b0da/ao0c02583_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/04a775e39fe7/ao0c02583_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/b2946dadb87b/ao0c02583_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/7a24d88ea4d2/ao0c02583_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/35a6753254e7/ao0c02583_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/4f6fa7ab6db8/ao0c02583_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/6c9db402c76a/ao0c02583_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/cbbe2838b0da/ao0c02583_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/04a775e39fe7/ao0c02583_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0bf8/7439397/b2946dadb87b/ao0c02583_0007.jpg

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