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开发一种基于生物传感器的免疫测定法以检测口腔癌患者漱口液中的人乳头瘤病毒E6癌蛋白。

Developing a Biosensor-Based Immunoassay to Detect HPV E6 Oncoprotein in the Saliva Rinse Fluid of Oral Cancer Patients.

作者信息

Cheng Chi-Sheng, Ou Bor-Rung, Lung Feng-Di

机构信息

Department of Chemistry, Tunghai University, Taichung 407224, Taiwan.

Department of Stomatology, Taichung Veterans General Hospital, Taichung 407219, Taiwan.

出版信息

J Pers Med. 2022 Apr 7;12(4):594. doi: 10.3390/jpm12040594.

DOI:10.3390/jpm12040594
PMID:35455710
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9027100/
Abstract

Transmission of Human papillomavirus (HPVs) is faithfully associated with carcinogenesis of oral cavity and oropharyngeal cancers. Therefore, clinical researchers may need to generate customized antibodies for the upcoming ELISA-based analysis to discover rare but valuable biomarkers. The aim of study was to develop and generate a biosensor-based immunoassay for early screening HPV-related oral cancer via saliva rinse fluid analysis. A peptide fragment of high-risk HPV subtype 16/18 protein, E6 protein (HP-1 protein sequence 48-66), was designed and synthesized, followed by the generation of polyclonal antibodies (anti-HP1 IgY) in our university-based laboratories. The titer and specificity of antibodies were determined by enzyme-linked immunosorbent assay (ELISA), and the Surface Plasmon Resonance (SPR) biosensor-based method was developed. Kinetic analyses by SPR confirmed that this designed peptide showed a high affinity with its generated polyclonal antibodies. Saliva fluid samples of thirty oral cancer patients and 13 healthy subjects were analyzed. SPR indicated that 26.8% of oral cancer patients had higher resonance unit (ΔRU) values than normal subjects. In conclusion, we developed a biosensor-based immunoassay to detect HPV E6 oncoprotein in the saliva rinse fluid for early screening and discrimination of HPV-related oral cancer patients.

摘要

人乳头瘤病毒（HPV）的传播与口腔癌和口咽癌的致癌作用密切相关。因此，临床研究人员可能需要为即将到来的基于酶联免疫吸附测定（ELISA）的分析生成定制抗体，以发现罕见但有价值的生物标志物。本研究的目的是通过漱口水分析开发并生成一种基于生物传感器的免疫测定方法，用于早期筛查HPV相关的口腔癌。设计并合成了高危HPV 16/18亚型E6蛋白的一个肽段（HP-1蛋白序列48-66），随后在我们的大学实验室中制备了多克隆抗体（抗-HP1 IgY）。通过酶联免疫吸附测定（ELISA）确定抗体的效价和特异性，并开发了基于表面等离子体共振（SPR）生物传感器的方法。SPR动力学分析证实，该设计肽与其产生的多克隆抗体具有高亲和力。分析了30例口腔癌患者和13名健康受试者的唾液样本。SPR结果表明，26.8%的口腔癌患者的共振单位（ΔRU）值高于正常受试者。总之，我们开发了一种基于生物传感器的免疫测定方法，用于检测漱口水样本中的HPV E6癌蛋白，以早期筛查和鉴别HPV相关的口腔癌患者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/452e33bff687/jpm-12-00594-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/5a09049e7331/jpm-12-00594-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/89b3b4e68cf0/jpm-12-00594-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/233675cc5b08/jpm-12-00594-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/7d408bb8ff31/jpm-12-00594-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/452e33bff687/jpm-12-00594-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/5a09049e7331/jpm-12-00594-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/89b3b4e68cf0/jpm-12-00594-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/233675cc5b08/jpm-12-00594-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/7d408bb8ff31/jpm-12-00594-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05e/9027100/452e33bff687/jpm-12-00594-g005.jpg

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