Magnetic multi-enzyme cascade combined with liquid chromatography tandem mass spectrometry for fast DNA digestion and quantitative analysis of 5-hydroxymethylcytosine in genome of human bladder cancer T24 cells induced by tetrachlorobenzoquinone
文献类型：期刊论文
| 作者 | Fang, Tianjiao ; Tang, Chao; Yin, Junfa; Wang, Hailin
|
| 刊名 | JOURNAL OF CHROMATOGRAPHY A
 |
| 出版日期 | 2022-08-02 |
| 卷号 | 1676期号:0页码:463279 |
| ISSN号 | 0021-9673 |
| 关键词 | DISINFECTION BY-PRODUCTS METAL-ORGANIC FRAMEWORKS BROMO-BENZOQUINONES ADDUCTS HALOBENZOQUINONES ENZYMES |
| 英文摘要 | Long-term exposure to halobenzoquinones (HBQs) can induce genomic damages and abnormal epige-netic modifications. High-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has shown unique advantages in identification and sensitive analysis of these structurally modified DNA lesions. Prior to MS analysis, genomic DNA needs to be fully digested into mono-nucleosides. Here, we prepared Supernuclease (SN)-, snake venom phosphodiesterase (SVP)-and calf intestinal alkaline phos-phatase (CIP)-individually immobilized magnetic nanoparticles (MNPs), and combined them according to certain formula to construct a recyclable SN-SVP-CIP magnetic nanoparticles (SNSC-MNPs) cascade for rapid and efficient DNA digestion. The SNSC-MNPs cascade can fully digest genomic DNA into mono -nucleosides within 30 min. The SNSC-MNPs cascade coupled with HPLC-MS/MS method can accurately and sensitively detect 5-hydroxymethylcytosine (5hmC) changes in genome of human bladder cancer T24 cells induced by tetrachlorobenzoquinone. The immobilization of enzymes on MNPs can enhance the sta-bility and enzymatic activity of the three enzymes, which guarantees the reusability and longtime preser-vation of the cascades. The relative digestive efficiencies are among 86%-106% up to ten times of reuse. The newly synthesized SNSC-MNPs cascade coupled with HPLC-MS/MS method is promising for fast iden-tification and analysis of structural modifications in genomic DNA. (c) 2022 Elsevier B.V. All rights reserved. |
| 源URL | [https://ir.rcees.ac.cn/handle/311016/47566]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 通讯作者 | Yin, Junfa |
| 作者单位 | 1.Chinese Acad Sci, Res Ctr Ecoenvironm Sci, State Key Lab Environm Chem & Ecotoxicol, Beijing 100085, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Jianghan Univ, Inst Environm & Hlth, Wuhan 430056, Peoples R China |
| 推荐引用方式 GB/T 7714 | Fang, Tianjiao,Tang, Chao,Yin, Junfa,et al. Magnetic multi-enzyme cascade combined with liquid chromatography tandem mass spectrometry for fast DNA digestion and quantitative analysis of 5-hydroxymethylcytosine in genome of human bladder cancer T24 cells induced by tetrachlorobenzoquinone[J]. JOURNAL OF CHROMATOGRAPHY A,2022,1676(0):463279. |
| APA | Fang, Tianjiao,Tang, Chao,Yin, Junfa,&Wang, Hailin.(2022).Magnetic multi-enzyme cascade combined with liquid chromatography tandem mass spectrometry for fast DNA digestion and quantitative analysis of 5-hydroxymethylcytosine in genome of human bladder cancer T24 cells induced by tetrachlorobenzoquinone.JOURNAL OF CHROMATOGRAPHY A,1676(0),463279. |
| MLA | Fang, Tianjiao,et al."Magnetic multi-enzyme cascade combined with liquid chromatography tandem mass spectrometry for fast DNA digestion and quantitative analysis of 5-hydroxymethylcytosine in genome of human bladder cancer T24 cells induced by tetrachlorobenzoquinone".JOURNAL OF CHROMATOGRAPHY A 1676.0(2022):463279. |
入库方式： OAI收割
来源：生态环境研究中心
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