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基于CD63核酸适配体功能化磁珠捕获外泌体，设计和制作了一种可从胃癌细胞上清液中分离出高纯度外泌体的微流控芯片，芯片内磁增强微结构的梳齿间距40μm,最小宽度仅为20μm。利用COMSOL软件对磁增强微结构进行磁场仿真，结果表明最大磁感应强度和磁场梯度分别可达15.1 T和5.77×104 T/m。通过纳米粒度Zeta电位仪对链霉亲和素磁珠与CD63核酸适配体偶联形成的功能化磁珠(SAMNP@Apt)进行表征，结果表明它们实现了良好偶联。使用SAMNP@Apt捕获外泌体，捕获效率可达92.7%。使用微电子机械系统(MEMS)技术制备了磁分离微流控芯片，并在微流控芯片中进行了外泌体磁分离实验，结果表明微流控芯片的外泌体分离纯度可达88.75%。
Abstract：Based on CD63-aptamer functionalized superparamagnetic beads, a microfluidic chip was designed and fabricated to separate high purity exosomes from gastric cancer cell supernatant. The magnetically enhanced microstructure in the chip has a comb pitch of 40 μm and a minimum width of only 20 μm. The magnetic field of the magnetic enhanced microstructure was simulated using COMSOL software. The results show that the maximum magnetic induction intensity and magnetic field gradient can reach 15.1 T and 5.77×104 T/m, respectively. The functionalized superparamagnetic beads(SAMNPs@Apt) were modified with CD63-aptamer by specific reaction between streptavidin on the surface of beads with biotin on the end of CD63-aptamers. The characterization result of the SAMNPs@Apt by nanoparticle Zeta potential analyzer shows that they achieve good coupling. The SAMNPs@Apt was used to capture exosomes with a capture efficiency of 92.7%. The magnetic separation microfluidic chip was fabricated by micro-electromechanical system(MEMS) technology, and the magnetic separation experiment of exosomes was performed in the microfluidic chip. The results show that the purity of exosome separation in the microfluidic chip can reach 88.75%.
[1] BRAY F,FERLAY J,SOERJOMATARAM I,et al.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries:global cancer statistics 2018 [J].CA:A Cancer Journal for Clinicians,2018,68(6):394-424.
[2] SMYTH E C,NILSSON M,GRABSCH H I,et al.Gastric cancer [J].Lancet,2020,396:635-648.
[3] LENGYEL C G,HUSSAIN S,TRAPANI D,et al.The emerging role of liquid biopsy in gastric cancer [J].Journal of Clinical Medicine,2021,10(10):2108-1-2108-16.
[4] DAASSI D,MAHONEY K M,FREEMAN G J.The importance of exosomal PDL1 intumour immune evasion [J].Nature Reviews Immunology,2020,20:209-215.
[5] PARK J,PARK J S,HUANG C H.An Integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma [J].Nature Biomedical Engineering,2021,5(7):678-689.
[6] SZSA B,FIA B.The Yin and Yang of exosome isolation methods:conventional practice,microfluidics,and commercial kits [J].Biotechnology Advances,2021,54(6):107814-1-107814-19.
[7] YANG D B,ZHANG W H,ZHANG H Y,et al.Progress,opportunity,and perspective on exosome isolation-efforts for efficient exosome-based theranostics [J].Theranostics,2020,10(8):3684-3707.
[8] B?RING A N,van der POL E,GROOTEMAAT A E,et al.Single-step isolation of extracellular vesicles from plasma by size-exclusion chromatography [J].Journal of Extracellular Vesicles,2014,3(1):23430-1-23430-12.
[9] VERGAUWEN G,DHONDT B,van DEUN J,et al.Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research [J].Scientific Reports,2017,7:2704-1-2704-12.
[10] ZHENG L,WANG H,ZUO P,et al.Rapid on-chip isolation of cancer-associated exosomes and combined analysis of exosomes and exosomal proteins [J].Analytical Chemistry,2022,94(21):7703-7712.
[11] SANCHO-ALBERO M,SEBASTIAN V,SESé J,et al.Isolation of exosomes from whole blood by a new microflui-dic device:proof of concept application in the diagnosis and monitoring of pancreatic cancer [J].Journal of Nanobiotechnology,2020,18(1):150-1-150-15.
[12] AYALA-MAR S,PEREZ-GONZALEZ V H,MATA-GóMEZ M A,et al.Electrokinetically driven exosome separation and concentration using dielectrophoretic-enhanced PDMS-based microfluidics [J].Analytical Chemistry,2019,91(23):14975-14982.
[13] QI R G,ZHU G X,WANG Y,et al.Microfluidic device for the analysis of MDR cancerous cell-derived exosomes' response tonanotherapy [J].Biomedical Microdevices,2019,21(2):35-1-35-9.
[14] KANG Y T,KIM Y J,BU J,et al.High-purity capture and release of circulating exosomes using an exosome-specific dual-patterned immunofiltration (ExoDIF) device [J].Nanoscale,2017,9(36):13495-13505.
[15] LIU C Y,SU C Q.Design strategies and application progress of therapeutic exosomes [J].Theranostics,2019,9(4):1015-1028.
[16] KANWAR S S,DUNLAY C J,SIMEONE D M,et al.Microfluidic device (ExoChip) for on-chip isolation,quantification and characterization of circulating exosomes [J].Lab on a Chip,2014,14(11):1891-1900.
[17] WANG J,LI W,ZHANG L C,et al.Chemically edited exosomes with dual ligand purified by microfluidic device for active targeted drug delivery to tumor cells [J].ACS Applied Materials & Interfaces,2017,9(33):27441-27452.
[18] WANG T,CHEN C Y,LARCHER L M,et al.Three decades of nucleic acidaptamer technologies:lessons learned,progress and opportunities on aptamer development [J].Biotechnology Advances.2019,37(1):28-50.
[19] WANG T,YIN W,ALSHAMAILEH H,et al.A detailed protein-SELEX protocol allowing visual assessments of individual steps for a high success rate [J].Human Gene Therapy Methods,2019,30(1):1-16.
[20] WANG T,RAHIMIZADEH K,VEEDU R N.Development of a novel DNA oligonucleotide targeting low-density lipoprotein receptor [J].Molecular Therapy-Nucleic Acids,2020,19:190-198.
[21] ZHANG K X,YUE Y L,WU S X,et al.Rapid capture and nondestructive release of extracellular vesicles using aptamer-based magnetic isolation [J].ACS Sensors,2019,4 (5):1245-1251.
[22] HEJAZIAN M,LI W H,NGUYEN N T.Lab on a chip for continuous-flow magnetic cell separation [J].Lab on a Chip,2015,15(4):959-970.
[23] FURLANI E P.Analysis of particle transport in amagnetophoretic microsystem [J].Journal of Applied Physics,2006,99(2):024912-1-024912-11.
基本信息:
DOI：10.13250/j.cnki.wndz.2023.05.009
中图分类号:TN492;R735.2
引用信息:
[1]王菲,林树靖,郅晓,等.基于MEMS技术的高纯度外泌体磁分离微流控芯片[J].微纳电子技术,2023,60(05):729-736.DOI:10.13250/j.cnki.wndz.2023.05.009.
基金信息:
国家自然科学基金青年科学基金项目(82003274); 上海市科委项目(21S31906400,21ZR1435000); 上海交通大学医工交叉项目(YG2022QN083)
2023-04-28
2023-04-28
2023-04-28