Automated determination of 8-OHdG in cells and tissue via immunofluorescence using a specially created antibody

Q1 Immunology and Microbiology Biotechnology Reports Pub Date : 2024-03-02 DOI:10.1016/j.btre.2024.e00833

Tobias Jung , Nicole Findik , Bianca Hartmann , Katja Hanack , Kai Grossmann , Dirk Roggenbuck , Marc Wegmann , René Mantke , Markus Deckert , Tilman Grune

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Abstract

Despite powerful DNA repair systems, oxidative damage/modification to DNA is an inevitable side effect of metabolism, ionizing radiation, lifestyle habits, inflammatory pathologies such as type-2 diabetes or metabolic syndrome, cancer and natural aging.

One of the most common oxidative DNA modifications is 8-OHdG (8‑hydroxy-2′-deoxyguanosine), which is the most widely used marker in research and clinical diagnostics. 8-OHdG is easily and specifically detectable in various samples such as urine, plasma, cells and tissues via a large variety of methods like ELISA, HPLC, chromatographic methods, and immunochemistry.

Formed by oxidation of guanine and being representative for the degree of DNA damage, 8-OHdG can be also used as biomarker for risk assessment of various cancers as well as degenerative diseases.

Here, we present a highly specific, self-developed 8-OHdG antibody in successful comparison to a commercially one, tested in cells (FF95, HCT116, and HT22) and intestinal tissue, focusing on automatized evaluation via fluorescence/confocal microscopy.

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使用特制抗体，通过免疫荧光自动测定细胞和组织中的 8-OHdG

尽管有强大的 DNA 修复系统，DNA 的氧化损伤/修饰仍是新陈代谢、电离辐射、生活习惯、炎症性病变（如 2 型糖尿病或代谢综合征）、癌症和自然衰老不可避免的副作用。8-OHdG（8-羟基-2′-脱氧鸟苷）是最常见的 DNA 氧化修饰之一，也是研究和临床诊断中最广泛使用的标记物。8-OHdG 可通过 ELISA、HPLC、色谱法和免疫化学等多种方法在尿液、血浆、细胞和组织等各种样本中轻松、特异地检测到。8-OHdG 由鸟嘌呤氧化形成，可代表 DNA 的损伤程度，也可用作各种癌症和退行性疾病风险评估的生物标记物。在这里，我们介绍了一种高度特异性的自主开发的 8-OHdG 抗体，并与市售抗体进行了成功对比，该抗体在细胞（FF95、HCT116 和 HT22）和肠道组织中进行了测试，重点是通过荧光/共聚焦显微镜进行自动评估。

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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来源期刊

Biotechnology Reports Immunology and Microbiology-Applied Microbiology and Biotechnology

CiteScore

15.80

自引率

0.00%

发文量

审稿时长

55 days

期刊介绍： Biotechnology Reports covers all aspects of Biotechnology particularly those reports that are useful and informative and that will be of value to other researchers in related fields. Biotechnology Reports loves ground breaking science, but will also accept good science that can be of use to the biotechnology community. The journal maintains a high quality peer review where submissions are considered on the basis of scientific validity and technical quality. Acceptable paper types are research articles (short or full communications), methods, mini-reviews, and commentaries in the following areas: Healthcare and pharmaceutical biotechnology Agricultural and food biotechnology Environmental biotechnology Molecular biology, cell and tissue engineering and synthetic biology Industrial biotechnology, biofuels and bioenergy Nanobiotechnology Bioinformatics & systems biology New processes and products in biotechnology, bioprocess engineering.