Ampholytic ion-exchange magnetic beads: a promising tool for selecting short fragments in circulating cell-free DNA analysis

作者全名：He, Gan; Wang, Weixuan; Zhou, Yongxia; Zhao, Guowei; Liao, Juan

作者地址：[He, Gan; Zhao, Guowei] Chongqing Med Univ, Yongchuan Hosp, Gastrointestinal Surg, Chongqing, Peoples R China; [Wang, Weixuan] Chongqing Med Univ, Inst Life Sci, Chongqing, Peoples R China; [Zhou, Yongxia] Chongqing Med Univ, Yongchuan Hosp, Dept Radiol, Chongqing, Peoples R China; [Zhou, Yongxia; Liao, Juan] Chongqing Med Univ, Yongchuan Hosp, Ctr Lab, Chongqing, Peoples R China; [Liao, Juan] Chongqing Geriatr Dis Clin Res Ctr, Chongqing, Peoples R China

通信作者：Liao, J (通讯作者)，Chongqing Med Univ, Yongchuan Hosp, Ctr Lab, Chongqing, Peoples R China.; Liao, J (通讯作者)，Chongqing Geriatr Dis Clin Res Ctr, Chongqing, Peoples R China.

来源：FRONTIERS IN ONCOLOGY

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:001228269100001

JCR分区：Q2

影响因子：3.5

年份：2024

卷号：14

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：ampholytic ion exchange; cell-free DNA; liquid biopsy; magnetic bead; magnetic submicron particle; silane; zwitterion

摘要：Objective For liquid biopsy of cancer, the extraction of circulating cell-free DNA (cfDNA) from plasma is required. We evaluated the efficacy of use of magnetic submicron particles coated with abundant small zwitterions (MSP-ZEWBs) for extracting short fragments of cfDNA.Methods We developed and optimized an MSP-ZEWB-based cfDNA extraction method using ampholytic ion-exchange materials and compared its results with those using a control kit. We measured the cfDNA concentration by quantitative polymerase-chain-reaction and using the Qubit method and analyzed cfDNA fragmentation patterns using a bioanalyzer.Results The fragment size of cfDNA isolated from glycine hydrochloric acid at a pH of 2.2 exhibited a better alignment with the DNA marker. The highest DNA intensity was observed at the final concentration of 0.8% polyethylene glycol 8000. The intensity of cfDNA decreased significantly when isolated from plasma with DNA marker using MSP-ZEWBs with an adsorption buffer containing guanidine hydrochloride or isothiocyanoguanidine. All fragments were successfully extracted using MSP-ZEWBs from both plasma and phosphate-buffered saline. Notably, the intensity of short cfDNA fragments isolated using MSP-ZEWBs remained consistent for recovery of long DNA fragments. indicating a potential selective of small fragments.Conclusion The extraction of plasma cfDNA with MSP-ZEWBs requires no protein denaturation, shows resistance to cells remaining in plasma, and demonstrates higher overall efficiency and better reproducibility than other extraction methods. Use of MSP-ZEWBs may greatly enhance liquid biopsy of cancers through the analysis of plasma cfDNA in clinical practice.

基金机构：National Nature Science Foundation of China [82102695]; Science and Health Joint Medical Research Project [2019QNXM013]; Chongqing Science and Technology Commission Project [CSTC2015JCYJA10043]; Natural Science Foundation Project of the Chongqing Education Committee [KJ1500239]

基金资助正文：The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was supported by the National Nature Science Foundation of China (No. 82102695), the Science and Health Joint Medical Research Project (2019QNXM013), the Chongqing Science and Technology Commission Project (CSTC2015JCYJA10043), and the Natural Science Foundation Project of the Chongqing Education Committee (KJ1500239).