表面修饰有阳离子的磁性二氧化硅微粒的制备及其应用
|
摘要
磁性复合微粒是超顺磁性纳米粒子与有机或无机材料通过包覆、交联等方式形成的纳微米级复合粒子。磁性复合微粒具有超顺磁性、较高的比表面积、可修饰功能基团等特点。因此,广泛应用于免疫学检测、核酸纯化、细胞分选、药物载体和酶的固定化等生物医学领域。本研究制备了两种表面修饰有阳离子的磁性二氧化硅微粒,基于两种磁性微粒建立了200μL人全血基因组DNA纯化方法,并对其进行优化。
采用微乳液法,以十六烷基三甲基溴化铵(CTAB)作为表面活性剂、甲苯为溶剂,与铁盐溶液(FeCl2和FeC13的混合溶液)剧烈搅拌形成微乳液,以氨水提供碱性环境合成Fe304磁性微粒,在原位以正硅酸乙酯(TEOS)提供硅源,直接在微乳液中包覆Fe304磁性微粒制备得到二氧化硅磁性微粒。再经修饰分别得到表面含有二乙氨基乙基(DEAE)和L-组氨酸(L-His)的两种磁性阴离子交换二氧化硅微粒。
基于表面修饰有二乙氨基乙基(DEAE)和L-组氨酸(L-His)的两种磁性阴离子交换二氧化硅微粒,以人全血作为样本,分别建立适用于200μL人全血基因组DNA纯化方法。并通过对纯化过程中裂解条件、结合条件、磁性微粒用量及清洗、洗脱条件等步骤对该方法进行优化,实验结果如下：
(1)对于200μL全血体系,DEAE-磁粒纯化可得到基因组DNA 3～5μg,其A260／A280值均介于1.70～1.90之间,纯化所得DNA分子结构完整,可用于PCR等分子生物学下游应用。但该方法需高盐洗脱,洗脱得到的DNA需经醇沉脱盐处理。
(2)基于His-磁粒200μL人全血基因组DNA纯化方法可纯化得到基因组DNA 3～4μg,其A260／A280值均介于1.65～1.8之间,相比前者纯化得到DNA的量较少,该方法结合所需pH5.0,可通过改变溶液的pH值洗脱,所需pH为8.5,洗脱得到的基因组DNA可直接用于下游分子生物学实验。
Magnetic particles is a kind of composite particles which composed of magnetite and some matrix such as organic or inorganic materials. Magnetic composite particles have the properties of superparamagnetism, surface effects and can be modified with functional groups. Therefore, they are widely used in research of immunology detection, nucleic purification, cell sorting, drug carrier and biological enzyme immobilization. Herein, two kinds of magnetic silica particles modified with cationic group (DEAE and His) were successfully prepared and used for genome DNA purification from 200μL human whole blood.
First, iron oxide nano-particles were synthesized by water-in-oil microemulsion technique, and in situ formed SiO2 as a shell on Fe3O4 particle surface. Based on core-shell structure magnetic silica nanoparticles (Fe3O4/SiO2), diethylaminoethyl (DEAE) and L-histidine (L-His) was modified on the particles and formed anion exchange magnetic silica particles respectively.
Using the particles modified with DEAE and L-His as carriers, genomic DNA was extracted from 200μL human whole blood, the steps including the lysis of whole blood, amount of the particles added, washing and eluting were optimized and the results show as follow:.
(1) 3-5μg genome DNA was obtained using 1.5 mg of DEAE-magnetic particles from 200μL human whole blood, the ratios of obtained genomic DNA were in the range of 1.70～1.90. However, the step of elution of this method need high salt concentration and need further ethanol precipitation to get genome DNA sample without salt.
(2) 3～4μg genome DNA was obtained using 1 mg of His-magnetic particles from 200μL human whole blood, the ratios of obtained genomic DNA were in the range of 1.65-1.80. The genome DNA yielded after the step of elution can be directly used in biological downstream applications. Because the elution step depends on the pH of the elution buffer using TE of pH 8.5.
采用微乳液法,以十六烷基三甲基溴化铵(CTAB)作为表面活性剂、甲苯为溶剂,与铁盐溶液(FeCl2和FeC13的混合溶液)剧烈搅拌形成微乳液,以氨水提供碱性环境合成Fe304磁性微粒,在原位以正硅酸乙酯(TEOS)提供硅源,直接在微乳液中包覆Fe304磁性微粒制备得到二氧化硅磁性微粒。再经修饰分别得到表面含有二乙氨基乙基(DEAE)和L-组氨酸(L-His)的两种磁性阴离子交换二氧化硅微粒。
基于表面修饰有二乙氨基乙基(DEAE)和L-组氨酸(L-His)的两种磁性阴离子交换二氧化硅微粒,以人全血作为样本,分别建立适用于200μL人全血基因组DNA纯化方法。并通过对纯化过程中裂解条件、结合条件、磁性微粒用量及清洗、洗脱条件等步骤对该方法进行优化,实验结果如下：
(1)对于200μL全血体系,DEAE-磁粒纯化可得到基因组DNA 3～5μg,其A260／A280值均介于1.70～1.90之间,纯化所得DNA分子结构完整,可用于PCR等分子生物学下游应用。但该方法需高盐洗脱,洗脱得到的DNA需经醇沉脱盐处理。
(2)基于His-磁粒200μL人全血基因组DNA纯化方法可纯化得到基因组DNA 3～4μg,其A260／A280值均介于1.65～1.8之间,相比前者纯化得到DNA的量较少,该方法结合所需pH5.0,可通过改变溶液的pH值洗脱,所需pH为8.5,洗脱得到的基因组DNA可直接用于下游分子生物学实验。
Magnetic particles is a kind of composite particles which composed of magnetite and some matrix such as organic or inorganic materials. Magnetic composite particles have the properties of superparamagnetism, surface effects and can be modified with functional groups. Therefore, they are widely used in research of immunology detection, nucleic purification, cell sorting, drug carrier and biological enzyme immobilization. Herein, two kinds of magnetic silica particles modified with cationic group (DEAE and His) were successfully prepared and used for genome DNA purification from 200μL human whole blood.
First, iron oxide nano-particles were synthesized by water-in-oil microemulsion technique, and in situ formed SiO2 as a shell on Fe3O4 particle surface. Based on core-shell structure magnetic silica nanoparticles (Fe3O4/SiO2), diethylaminoethyl (DEAE) and L-histidine (L-His) was modified on the particles and formed anion exchange magnetic silica particles respectively.
Using the particles modified with DEAE and L-His as carriers, genomic DNA was extracted from 200μL human whole blood, the steps including the lysis of whole blood, amount of the particles added, washing and eluting were optimized and the results show as follow:.
(1) 3-5μg genome DNA was obtained using 1.5 mg of DEAE-magnetic particles from 200μL human whole blood, the ratios of obtained genomic DNA were in the range of 1.70～1.90. However, the step of elution of this method need high salt concentration and need further ethanol precipitation to get genome DNA sample without salt.
(2) 3～4μg genome DNA was obtained using 1 mg of His-magnetic particles from 200μL human whole blood, the ratios of obtained genomic DNA were in the range of 1.65-1.80. The genome DNA yielded after the step of elution can be directly used in biological downstream applications. Because the elution step depends on the pH of the elution buffer using TE of pH 8.5.
引文
[1]Jun Hong, Dongmei Xu, Peijun Gong. Covalent binding of a-chymotrypsin on the magnetic nanogels covered by amino group[J]. Journal of Molecular Catalysis B: Enzymatic,2007,45:84-90
[2]Victor N. Morozov, Tamara Y. Morozova. Active bead-linked immunoassay on protein microarry[J]. Analytica Chimica Acta,2006,564:40-52
[3]Lu A. H., Schmidt W., Matoussevitch N., et al. Nanoengineering of a magnetically separable hydrogenation catalyst[J]. Angewandte Chemie International Edition,2004,43: 4304-4306
[4]Tsang S. C., Caps V., Paraskevas I., et al. Magneticaly separable, caron-supported nanocatalysts for the manufacture of fine chemicals[J]. Angewandte Chemie International Edition,2004,439(42):5645-5645
[5]Gupta A. K., Gupta M. Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications[J]. Biomaterials,2005,26(18):3995-4021
[6]Li Z., Wei L., Gao M. Y, et al. One-Pot Reaction to Synthesize Biocompatible Magnetite Nanoparticles[J]. Advanced Materials,2005,17(8):1001-1005
[7]Neveu S., Bee A., Robineau M., et al. Size-selective chemical synthesis of tartrate stabilized cobalt ferrite ionic magnetic fluid[J]. Joural of Colloid and Interface Science, 2002,255(2):294-298
[8]Grasset F., Labhsetwar N., Li D., et al. Mimicking the Lotus Effect:Influence of Double Roughness Structrures and Slender Pillars[J]. Langmuir,2002,18(21):8209-8216
[9]Sum S., Zeng H. Size-controlled synthesis of magnetite nanoparticles[J]. Journal of the American Chemical Society,2002,124(28):8204-8205
[10]Chen Q., Rondinone A. J., Chakoumakos B. C., et al. Synthesis of superparamagnetic MgFe2O4 nanoparticles by corecipitation[J]. Journal of Magnetism and Magnetic Materials, 1994,194:1-7
[11]Park J., An K., Hwang Y, et al. Ultra-large-scale syntheses of monAisperse
nanocrystals[J]. Nature Material,2004,3(28):891-895
[12]Sun S., Murray C. B., Weller D., et al. MonAisperse FePt Nanoparticles and
Ferromagnetic Fept Nanocrystal Srperlattices[J]. Science,2000,287:1989-1992
[13]Shevchenko E. V., Rogach A. L., et al. Colloidal Synthese and Self-Assembly of CoPt3Nanocrystals[J]. Journal of American Chemical Society,2002,124(38):11480
[14]郑兰香,彭国新.超细四氧化三铁微粒的制备[J].精细化工,1995,12(6)：11-13
[15]陈辉.高温分解法合成Fe304磁性纳米微粒[J].河南化工,2004,2：11-12
[16]Jana N. R., Chen Y., Peng X. Size and Shape-Controlled Magnetic (Cr,Mn,Fe,Co,Ni) Oxide Nanocrystals via a Simple and General Approach[J]. Chemistry of Materials 2004, 16(20):3931-3935
[17]Xu Jing, Yang Haibin, Fu Wuyou. Preparation and magnetic properties of magnetic nanparticles by sol-gel method[J]. J. Magn. Magn. Mater.,2007,309(1):307-311
[18]Koneracka M., Kopcansky P., Timkom, et al. Direet binding procedure of proteins and enzynles to fine magnetie partieles[J]. Journal of Magnetism and Magnetie Materials,2002, 252:409-411
[19]S.H. Huang, M.H. Liao, D.H. Chen. Direct binding and eharaeterization of lipase onto magnetie nanoparticles[J]. Biotechonoloy Progress,2003,19:1095-1100
[20]C. Bergemann, Muller-Schulte D., Oster J., et al. Magnetic ion-exchange nano-and microparticles for medical, biochemical and molecular biological application[J]. Journal Magn Mater,1999,194(1-3):45
[21]Dekker R. F. M. Immobilization of alactase onto a magnetic support by covalent attachment to polyethyleneimine-gluta-raldehyde-actibated magnetite[J]. Appl Biochem Biotechn,1989,22:289
[22]Cuyper M. D., Jonian M. Mechanistic aspects of the adsorption of phsopho lipids onto lauticacid stabilized magnetite nanocolloids[J]. Langmuir,1990,7:647
[23]Cooker T. M., Fee C. J., Evans R. A. Preparation of magnetically susceptible plyacrylamide/magnetite beads for use in magnetically stabilized fluidezed bed chromatography[J]. Biotechn Bioeng,1997,53(1):79
[24]Akkaya B., Uzun L., Candan F, et al. N-methacryloyl-(L)-histidine methyl ester carrying porous magnetic beads for metal chelate adsorption of cytochromec[J]. Mater Sci Eng:C, 2007,27(1):180
[25]Gan L. M., Chan H. S., Chan H. S. Preparation of conducting polyaniline-couated barium sulfate nanoparticles in inverse microemulsions[J]. Mater Chem Phys,1995,40(2):94
[26]Kawaguchi H. Functional polymer microspheres[J]. Prog Polym Sci,2000,25(8):1171
[27]Horak D, Semenyuk N., Lednieky F. Effect of the reaction parameters on the particle size in the dispersion polymerization of 2-hydroxyethyl and glycidyl methacrylate in the presence of a ferrofluid[J]. J Polym Sci:Part A:Polym Chem,2003,41(12):1848
[28]Iwamura I. H., Sugawara T., Itoh K., et al. High-spin polycarbenes as amodel for orgnic ferromagnets[J]. Mol Cyst Liq Cryst,1985,125:379
[29]Ma Z. Y., Guan Y. P., Liu H. Z. Synthesis and characterization of micron-sized monodisperse superparamagnetic polymer partclesw with amino groups[J]. J Polym Sci: Part A:Polym Chem,2005,43:3433
[30]Santra S., Tapee R., Theodoro Poulou N., et al. Synthesis and characterization of silica-coated iron oxide nanoparticles in microemusion:The effect of nonionic surfactants[J]. Langmuir,2001,17:2900
[31]Hafeli U. O., Sweeney S. M., Beresford B. A., et al. Effective targeting of magnetic radioactive 90 Y-microspheres to tumor cells by an externally applied magnetic field[J]. Nuclear Medicine and Biology,1995,22(2):147
[32]Denkbas E. B., Kilicay E., Birliksenven C., et al. Magnetic chitosan microspheres: preparation and characterization[J]. React Funct Polym,2002,50(3):225
[33]Caruso F., Susha A. S., Giersig M., et al. Magnetic core-shell particles:Preparation of magnetite multilayers on polymer latex microsheres[J]. Adv Mater,1999,11:950
[34]Chen S. A., Chang H. S. Kinetics and mechanism of emulsifier free emulsion polymeriztion III Styene/nonionic comonmer(2-hydroxyethyl methacrylate) system[J]. J Polym Sci:Part A:Polym Chem,1990,28(9):2547
[35]Furusawa K., Nagashima K. Synthetic process to control the total size and component distribution of multilayer magnetic composite particles[J]. Collid Polym Sci,1994, 272(1104):1104-1110
[36]Caruso F., Caruso R. A., Mohwald H. Nanoengineering of inorganic and hybrid hollow spheres by colloidal templating[J]. Science,1998,282(6):1111
[37]Zhu Y., Hong D., et al. Preparation and characterization of core-shell monodisperesed magnetic silica microshperes[J]. Collides Surf A:Physicochemical and Enginerring Aspects, 2003,231:123
[38]Garcaa A. A., Bonen M. R., Ramirez V. J., et al. Bioseparation Process Science[J]. Tsinghua University Academic Press,2002:229-314
[39]Alexiou C., Schmidt A., Klein R., et al. Magnetic drug targeting:bio-distribution and dependency on magnetic field strength[J]. J. Magn. Magn. Mater.,2002,252:363-366
[40]Jordan A., Scholz R., Maier Hauff K., et al. Presentation of a new magnetic field therapy system for the treatment of human solid turners with magnetic fluid hyperthermia[J]. J. Magn. Magn. Mater.,2001,225:118-126
[41]Dailey J.P., Phillips J.P., Li C., et al. Synthesis of silicone magnetic fluid for use in eye surgery[J]. J. Magn. Magn. Mater.,1999,194:140-148
[42]Yoshimoto T., Mihama T., Takahashi K., et al. Chemical modification of enzymes with activated magnetic modifier[J]. Biochemical and Biophysical Research Communications, 1987,145(2):908-914
[43]Wang Qiu-Yu, Qin Chuan-Guang, Zhang Qiu-Yu, et al. Immobilization of papain by layer-by-layer self-assembly on magnetic polystyrene microspheres[J]. Chinese Journal of Applied Chemistry,2009,26(5):557-561
[44]Liu Li-Ping, Wu Ze-Zhi, Li Ping, et al. Recent advances of magnetic natural polymer microspheres as vehicles for targeted delivery [J]. Pharmaceutical Biotechnology,2004, 11(1):68-70
[45]Rittich B., Spanova A., Ohlashennyy Y. Characterization of deoxyribonuclease I immobilized on magnetic hydrophilic polymer particles[J]. J Chromatogr B,2002,774(1): 25-31
[46]Richardson J., Hawkins P., Luxton R. The use of coated paramagnetic particles as a physical label in a magneto-immunoassay[J]. Biosensors&Bioelectronics,2001,16: 989-993
[47]Schoepf U., Marecos E., Jain R., et al. Magnetic Labeling of Lymphocytes for in vivo Trafficking Studies[J]. Biotechniques,1998,24:642-651
[48]Handgretinger R., Lang P., Schumm M., et al. Isolation and transplantation of autologous peripheral CD34+ progenitor cells highly purified by magnetic-activated sorting[J]. Bone Marrow Transplant,1998,21:987-993
[49]Gupta A. K., Gupta M. Synthesis and surface engineering of iron oxide nanoparticles for biomedical application[J]. Biomaterials,2005,26:3995-4021
[50]Safarik I., Safarikova M. Use of magnetic techniques for the isolation of cells[J]. J. Chromatogr B Biomed Sci Appli,1999,722:33-53
[51]Tartaj P., Morales M. P., Gonzalez C. T., et al. Advaneed in magnetic nanoparticles for bioteehnology applications[J]. Journal of Magnetism and Magnetie Materials,2005, 290-291:28-34
[52]Bautista M. C., Bomati M. O., Zhang X., et al. Comparative study of ferrofluids based on dextran coated iron oxide and metal nanopartieles for contrast agents in magnetic resonane eimaging[J]. Nanoteehnology,2004,15:154-159
[53]Horak D., Bohhek J., Lubrt J. Magnetic poly (-hydroxyethyl methacrylate-co-ethylene dim ethacrylate) microspheres by dispersion ploymerzation[J]. Journal of Polymer Science Part A:Polymer Chemistry,2000,38:1161-1171
[54]Lafarik I., Lafarikova M. Magnetic nanoparticles and biosciences[J]. Monatshefte fur Chemie,2002,133:746-754
[55]Yuan J. J., Annes S. P., Takabayashi Y., et al. Synthesis of Biocompatible Poly[2-(methaeryloxy) ethyl phosphorylcholine]-Coated Magnetite Nanopartieles[J]. Langnluir,2006,22(26):10989-10993
[56]景猛,刘新权,刘恩重.神经干细胞超顺磁性氧化铁纳米粒子标记的体内MRI失踪[J].中华神经外科疾病研究杂志,2006,5(1)：92-93
[57]John Wilcockson. The Use of Sodium Perchlorate in Deproteinization during the Preparation of Nucleic Acids[J]. Biochem. J.,1973,135:559-561
[58]Bowtell Dd. Rapid isolation of eukaryotic DNA[J]. Anal.Biochem.,1987,162:463-465
[59]裴杰萍,端青.DNA提取方法的研究进展[J].微生物学免疫学进展,2004,32(3)：76-78
[60]Vogelstein B, Gillespie D. Preparative and analytical purification of DNA from agarose[J]. Proc. Natl. Acad. Sci.,1979,76:615-619
[61]Merzak Ka, Sherwood Cs, Turner Rfb, et al. Driving forces for DNA adsorption to silica in perchlorate solutions[J]. J. Colloic Interface Sci.,1996,181:635-644
[62]Boom R, Sol Cj, Salimans Mm, et al. Rapid and simple method for purification of nucleic acids[J]. J. Clin. Microbiol.,1990,28:495-503
[63]Boom R., Sol C., Beld M., et al. Improved silica-guanidinium thiocyanate DNA isolation procedure based on selective binding of bovine alpha-casein to silica particles[J]. J. Clin. Microbiol.,1999,37:615-619
[64]Breadmore Mc, Wolfe Ka, Arcbal Ig, et al. Microchip-based purification of DNA from biological samples[J]. Anal. Chem.,2003,75(8):1880-1886
[65]Yoshio Katoa, Masaaki Sasakia, Tsutomu Hashimotoa, et al. Separation of DNA restriction fragments by high-performance ion-exchange chromatography [J]. Journal of Chromatography A 1983,263:342-346
[66]Wahh P., Metzger D. A. Higuchi R.,Chdex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic Materil[J]. Biotechniques,1991,10:506-513
[67]Larnballerie X. D., Zandoui C. A.. One-step microbial DNA extraction method using "Chelex 100" suitable for gene amplification[J]. Res Micrebiol 1992,1992,143:758-790
[68]Erb W., Dobler I. W. Detection polyehlorinated biphenyl genes in polluted sediments by direct DNA extraction and polymerase chain reaction[J]. Appl. Environ. Microb.,1993,59: 4065-4073
[69]Romanowski G., Lorenz M.G., Wackernagel W. Adsorption of plasmid DNA to mineral surfaces and protection against DNase I [J]. Appl. Environ. Microb.,1991,57(4): 1056-1061
[70]Hawkinst. L., O'connor-Morin T., Roy A. DNA purification and isolation using a solid-Phase[J]. Nueleie Aeids Reseaeh,1994,22(21):4544-4544
[71]Peter R. L., Stephen E. B., Jon D. Recent developments of magnetie beads for use in nueleic acid purification[J]. Joumal of Chromatography A,1998,816:107-111
[72]Davies M. J., Taylor J. I., Sachsinger N. Isolation of plasmid DNA using magnetite as a solid-phase adsorbent[J]. Anal. Biochem.,1998,262:92-94
[73]Taylor J. I., Hurst C. D., Davies M. J. Application of magnetite and silica-magnetite composites to the isolation of genomic DNA[J]. Journal of Chromatography A,2000, 890(1):159-166
[74]Jurgen O., Jeffrey P., Lothara B. Polyvinyl-alchol-Eased magnetic beads for rapid an efficient separation of specific or unspecific nucleic acid sequences[J]. Journal of Magnetism and Magnetic Materials,2001,225:145
[75]Elkin C. J., Richardson P. M., Fourcade H. M. High throughput plasmid purifieation for capillary sequeneing[J]. Genome Research,2001,11(7):1269-1274
[76]Bach H. J., Hartmann A., Trevors J. T., et al. Magneti capture-hybridization method for
purifieation and probing of mRNA for neutral protease of Bacillus cereus[J]. Journal of Microbiological Methods,1999,37:173-187
[77]Zhao X., Rovelyn T. D., Wang K. Collection of trace amounisof DNA/mRNA molecules using genomagnetic nanocapturers[J]. Analytical Chemistry,2003,75(14):3476-3483
[78]Levison Peter R., Badger Stephen E., Dennis Jon, et al. Recent developments of magnetic beads for use in nucleic acid purification[J]. Journal of Chromatography A,1998, 816:107-111
[79]Levison Peter R., Badger Stephen E., Dennis Jon, et al. New approaches to the isolation of DNA by ion-exchange chromatography[J]. Journal of Chromatography A,1998,827: 337-344
[80]Brandon Yoza, Mitsufumi Matsumoto, Tadashi Matsunaga. DNA extraction using modified bacterial magnetic particles in the presence of amino silane compound[J]. jounal of Biotechnology,2001,94:217-224
[81]Brandon Yoza, Atsushi Arakaki, Tadashi Matsunaga. DNA extraction using bacterial magnetic particles modified with hyperbranched polyamidoamine dendrimer[J]. Journal of Biotechnology,2002,101:219-228
[82]孙敏莉,王强斌,张皓,et al. DEAE-磁粒纳米粒子提纯质粒的研究[J].上海第二医科大学学报,2003,23(6)：512-514
[83]Chen-Li Chiang, Ching-Shan Sung, Ting-Feng Wu, et al. Application of superparamagnetic nanoparticles in purification of plasmid DNA from bacterial cells[J]. Journal of Chromatography B,2005,822:54-60
[84]Matthew John Baker. Isolation of nucleic acids[P]. US patent:2007/0231892,2007-10-4
[2]Victor N. Morozov, Tamara Y. Morozova. Active bead-linked immunoassay on protein microarry[J]. Analytica Chimica Acta,2006,564:40-52
[3]Lu A. H., Schmidt W., Matoussevitch N., et al. Nanoengineering of a magnetically separable hydrogenation catalyst[J]. Angewandte Chemie International Edition,2004,43: 4304-4306
[4]Tsang S. C., Caps V., Paraskevas I., et al. Magneticaly separable, caron-supported nanocatalysts for the manufacture of fine chemicals[J]. Angewandte Chemie International Edition,2004,439(42):5645-5645
[5]Gupta A. K., Gupta M. Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications[J]. Biomaterials,2005,26(18):3995-4021
[6]Li Z., Wei L., Gao M. Y, et al. One-Pot Reaction to Synthesize Biocompatible Magnetite Nanoparticles[J]. Advanced Materials,2005,17(8):1001-1005
[7]Neveu S., Bee A., Robineau M., et al. Size-selective chemical synthesis of tartrate stabilized cobalt ferrite ionic magnetic fluid[J]. Joural of Colloid and Interface Science, 2002,255(2):294-298
[8]Grasset F., Labhsetwar N., Li D., et al. Mimicking the Lotus Effect:Influence of Double Roughness Structrures and Slender Pillars[J]. Langmuir,2002,18(21):8209-8216
[9]Sum S., Zeng H. Size-controlled synthesis of magnetite nanoparticles[J]. Journal of the American Chemical Society,2002,124(28):8204-8205
[10]Chen Q., Rondinone A. J., Chakoumakos B. C., et al. Synthesis of superparamagnetic MgFe2O4 nanoparticles by corecipitation[J]. Journal of Magnetism and Magnetic Materials, 1994,194:1-7
[11]Park J., An K., Hwang Y, et al. Ultra-large-scale syntheses of monAisperse
nanocrystals[J]. Nature Material,2004,3(28):891-895
[12]Sun S., Murray C. B., Weller D., et al. MonAisperse FePt Nanoparticles and
Ferromagnetic Fept Nanocrystal Srperlattices[J]. Science,2000,287:1989-1992
[13]Shevchenko E. V., Rogach A. L., et al. Colloidal Synthese and Self-Assembly of CoPt3Nanocrystals[J]. Journal of American Chemical Society,2002,124(38):11480
[14]郑兰香,彭国新.超细四氧化三铁微粒的制备[J].精细化工,1995,12(6)：11-13
[15]陈辉.高温分解法合成Fe304磁性纳米微粒[J].河南化工,2004,2：11-12
[16]Jana N. R., Chen Y., Peng X. Size and Shape-Controlled Magnetic (Cr,Mn,Fe,Co,Ni) Oxide Nanocrystals via a Simple and General Approach[J]. Chemistry of Materials 2004, 16(20):3931-3935
[17]Xu Jing, Yang Haibin, Fu Wuyou. Preparation and magnetic properties of magnetic nanparticles by sol-gel method[J]. J. Magn. Magn. Mater.,2007,309(1):307-311
[18]Koneracka M., Kopcansky P., Timkom, et al. Direet binding procedure of proteins and enzynles to fine magnetie partieles[J]. Journal of Magnetism and Magnetie Materials,2002, 252:409-411
[19]S.H. Huang, M.H. Liao, D.H. Chen. Direct binding and eharaeterization of lipase onto magnetie nanoparticles[J]. Biotechonoloy Progress,2003,19:1095-1100
[20]C. Bergemann, Muller-Schulte D., Oster J., et al. Magnetic ion-exchange nano-and microparticles for medical, biochemical and molecular biological application[J]. Journal Magn Mater,1999,194(1-3):45
[21]Dekker R. F. M. Immobilization of alactase onto a magnetic support by covalent attachment to polyethyleneimine-gluta-raldehyde-actibated magnetite[J]. Appl Biochem Biotechn,1989,22:289
[22]Cuyper M. D., Jonian M. Mechanistic aspects of the adsorption of phsopho lipids onto lauticacid stabilized magnetite nanocolloids[J]. Langmuir,1990,7:647
[23]Cooker T. M., Fee C. J., Evans R. A. Preparation of magnetically susceptible plyacrylamide/magnetite beads for use in magnetically stabilized fluidezed bed chromatography[J]. Biotechn Bioeng,1997,53(1):79
[24]Akkaya B., Uzun L., Candan F, et al. N-methacryloyl-(L)-histidine methyl ester carrying porous magnetic beads for metal chelate adsorption of cytochromec[J]. Mater Sci Eng:C, 2007,27(1):180
[25]Gan L. M., Chan H. S., Chan H. S. Preparation of conducting polyaniline-couated barium sulfate nanoparticles in inverse microemulsions[J]. Mater Chem Phys,1995,40(2):94
[26]Kawaguchi H. Functional polymer microspheres[J]. Prog Polym Sci,2000,25(8):1171
[27]Horak D, Semenyuk N., Lednieky F. Effect of the reaction parameters on the particle size in the dispersion polymerization of 2-hydroxyethyl and glycidyl methacrylate in the presence of a ferrofluid[J]. J Polym Sci:Part A:Polym Chem,2003,41(12):1848
[28]Iwamura I. H., Sugawara T., Itoh K., et al. High-spin polycarbenes as amodel for orgnic ferromagnets[J]. Mol Cyst Liq Cryst,1985,125:379
[29]Ma Z. Y., Guan Y. P., Liu H. Z. Synthesis and characterization of micron-sized monodisperse superparamagnetic polymer partclesw with amino groups[J]. J Polym Sci: Part A:Polym Chem,2005,43:3433
[30]Santra S., Tapee R., Theodoro Poulou N., et al. Synthesis and characterization of silica-coated iron oxide nanoparticles in microemusion:The effect of nonionic surfactants[J]. Langmuir,2001,17:2900
[31]Hafeli U. O., Sweeney S. M., Beresford B. A., et al. Effective targeting of magnetic radioactive 90 Y-microspheres to tumor cells by an externally applied magnetic field[J]. Nuclear Medicine and Biology,1995,22(2):147
[32]Denkbas E. B., Kilicay E., Birliksenven C., et al. Magnetic chitosan microspheres: preparation and characterization[J]. React Funct Polym,2002,50(3):225
[33]Caruso F., Susha A. S., Giersig M., et al. Magnetic core-shell particles:Preparation of magnetite multilayers on polymer latex microsheres[J]. Adv Mater,1999,11:950
[34]Chen S. A., Chang H. S. Kinetics and mechanism of emulsifier free emulsion polymeriztion III Styene/nonionic comonmer(2-hydroxyethyl methacrylate) system[J]. J Polym Sci:Part A:Polym Chem,1990,28(9):2547
[35]Furusawa K., Nagashima K. Synthetic process to control the total size and component distribution of multilayer magnetic composite particles[J]. Collid Polym Sci,1994, 272(1104):1104-1110
[36]Caruso F., Caruso R. A., Mohwald H. Nanoengineering of inorganic and hybrid hollow spheres by colloidal templating[J]. Science,1998,282(6):1111
[37]Zhu Y., Hong D., et al. Preparation and characterization of core-shell monodisperesed magnetic silica microshperes[J]. Collides Surf A:Physicochemical and Enginerring Aspects, 2003,231:123
[38]Garcaa A. A., Bonen M. R., Ramirez V. J., et al. Bioseparation Process Science[J]. Tsinghua University Academic Press,2002:229-314
[39]Alexiou C., Schmidt A., Klein R., et al. Magnetic drug targeting:bio-distribution and dependency on magnetic field strength[J]. J. Magn. Magn. Mater.,2002,252:363-366
[40]Jordan A., Scholz R., Maier Hauff K., et al. Presentation of a new magnetic field therapy system for the treatment of human solid turners with magnetic fluid hyperthermia[J]. J. Magn. Magn. Mater.,2001,225:118-126
[41]Dailey J.P., Phillips J.P., Li C., et al. Synthesis of silicone magnetic fluid for use in eye surgery[J]. J. Magn. Magn. Mater.,1999,194:140-148
[42]Yoshimoto T., Mihama T., Takahashi K., et al. Chemical modification of enzymes with activated magnetic modifier[J]. Biochemical and Biophysical Research Communications, 1987,145(2):908-914
[43]Wang Qiu-Yu, Qin Chuan-Guang, Zhang Qiu-Yu, et al. Immobilization of papain by layer-by-layer self-assembly on magnetic polystyrene microspheres[J]. Chinese Journal of Applied Chemistry,2009,26(5):557-561
[44]Liu Li-Ping, Wu Ze-Zhi, Li Ping, et al. Recent advances of magnetic natural polymer microspheres as vehicles for targeted delivery [J]. Pharmaceutical Biotechnology,2004, 11(1):68-70
[45]Rittich B., Spanova A., Ohlashennyy Y. Characterization of deoxyribonuclease I immobilized on magnetic hydrophilic polymer particles[J]. J Chromatogr B,2002,774(1): 25-31
[46]Richardson J., Hawkins P., Luxton R. The use of coated paramagnetic particles as a physical label in a magneto-immunoassay[J]. Biosensors&Bioelectronics,2001,16: 989-993
[47]Schoepf U., Marecos E., Jain R., et al. Magnetic Labeling of Lymphocytes for in vivo Trafficking Studies[J]. Biotechniques,1998,24:642-651
[48]Handgretinger R., Lang P., Schumm M., et al. Isolation and transplantation of autologous peripheral CD34+ progenitor cells highly purified by magnetic-activated sorting[J]. Bone Marrow Transplant,1998,21:987-993
[49]Gupta A. K., Gupta M. Synthesis and surface engineering of iron oxide nanoparticles for biomedical application[J]. Biomaterials,2005,26:3995-4021
[50]Safarik I., Safarikova M. Use of magnetic techniques for the isolation of cells[J]. J. Chromatogr B Biomed Sci Appli,1999,722:33-53
[51]Tartaj P., Morales M. P., Gonzalez C. T., et al. Advaneed in magnetic nanoparticles for bioteehnology applications[J]. Journal of Magnetism and Magnetie Materials,2005, 290-291:28-34
[52]Bautista M. C., Bomati M. O., Zhang X., et al. Comparative study of ferrofluids based on dextran coated iron oxide and metal nanopartieles for contrast agents in magnetic resonane eimaging[J]. Nanoteehnology,2004,15:154-159
[53]Horak D., Bohhek J., Lubrt J. Magnetic poly (-hydroxyethyl methacrylate-co-ethylene dim ethacrylate) microspheres by dispersion ploymerzation[J]. Journal of Polymer Science Part A:Polymer Chemistry,2000,38:1161-1171
[54]Lafarik I., Lafarikova M. Magnetic nanoparticles and biosciences[J]. Monatshefte fur Chemie,2002,133:746-754
[55]Yuan J. J., Annes S. P., Takabayashi Y., et al. Synthesis of Biocompatible Poly[2-(methaeryloxy) ethyl phosphorylcholine]-Coated Magnetite Nanopartieles[J]. Langnluir,2006,22(26):10989-10993
[56]景猛,刘新权,刘恩重.神经干细胞超顺磁性氧化铁纳米粒子标记的体内MRI失踪[J].中华神经外科疾病研究杂志,2006,5(1)：92-93
[57]John Wilcockson. The Use of Sodium Perchlorate in Deproteinization during the Preparation of Nucleic Acids[J]. Biochem. J.,1973,135:559-561
[58]Bowtell Dd. Rapid isolation of eukaryotic DNA[J]. Anal.Biochem.,1987,162:463-465
[59]裴杰萍,端青.DNA提取方法的研究进展[J].微生物学免疫学进展,2004,32(3)：76-78
[60]Vogelstein B, Gillespie D. Preparative and analytical purification of DNA from agarose[J]. Proc. Natl. Acad. Sci.,1979,76:615-619
[61]Merzak Ka, Sherwood Cs, Turner Rfb, et al. Driving forces for DNA adsorption to silica in perchlorate solutions[J]. J. Colloic Interface Sci.,1996,181:635-644
[62]Boom R, Sol Cj, Salimans Mm, et al. Rapid and simple method for purification of nucleic acids[J]. J. Clin. Microbiol.,1990,28:495-503
[63]Boom R., Sol C., Beld M., et al. Improved silica-guanidinium thiocyanate DNA isolation procedure based on selective binding of bovine alpha-casein to silica particles[J]. J. Clin. Microbiol.,1999,37:615-619
[64]Breadmore Mc, Wolfe Ka, Arcbal Ig, et al. Microchip-based purification of DNA from biological samples[J]. Anal. Chem.,2003,75(8):1880-1886
[65]Yoshio Katoa, Masaaki Sasakia, Tsutomu Hashimotoa, et al. Separation of DNA restriction fragments by high-performance ion-exchange chromatography [J]. Journal of Chromatography A 1983,263:342-346
[66]Wahh P., Metzger D. A. Higuchi R.,Chdex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic Materil[J]. Biotechniques,1991,10:506-513
[67]Larnballerie X. D., Zandoui C. A.. One-step microbial DNA extraction method using "Chelex 100" suitable for gene amplification[J]. Res Micrebiol 1992,1992,143:758-790
[68]Erb W., Dobler I. W. Detection polyehlorinated biphenyl genes in polluted sediments by direct DNA extraction and polymerase chain reaction[J]. Appl. Environ. Microb.,1993,59: 4065-4073
[69]Romanowski G., Lorenz M.G., Wackernagel W. Adsorption of plasmid DNA to mineral surfaces and protection against DNase I [J]. Appl. Environ. Microb.,1991,57(4): 1056-1061
[70]Hawkinst. L., O'connor-Morin T., Roy A. DNA purification and isolation using a solid-Phase[J]. Nueleie Aeids Reseaeh,1994,22(21):4544-4544
[71]Peter R. L., Stephen E. B., Jon D. Recent developments of magnetie beads for use in nueleic acid purification[J]. Joumal of Chromatography A,1998,816:107-111
[72]Davies M. J., Taylor J. I., Sachsinger N. Isolation of plasmid DNA using magnetite as a solid-phase adsorbent[J]. Anal. Biochem.,1998,262:92-94
[73]Taylor J. I., Hurst C. D., Davies M. J. Application of magnetite and silica-magnetite composites to the isolation of genomic DNA[J]. Journal of Chromatography A,2000, 890(1):159-166
[74]Jurgen O., Jeffrey P., Lothara B. Polyvinyl-alchol-Eased magnetic beads for rapid an efficient separation of specific or unspecific nucleic acid sequences[J]. Journal of Magnetism and Magnetic Materials,2001,225:145
[75]Elkin C. J., Richardson P. M., Fourcade H. M. High throughput plasmid purifieation for capillary sequeneing[J]. Genome Research,2001,11(7):1269-1274
[76]Bach H. J., Hartmann A., Trevors J. T., et al. Magneti capture-hybridization method for
purifieation and probing of mRNA for neutral protease of Bacillus cereus[J]. Journal of Microbiological Methods,1999,37:173-187
[77]Zhao X., Rovelyn T. D., Wang K. Collection of trace amounisof DNA/mRNA molecules using genomagnetic nanocapturers[J]. Analytical Chemistry,2003,75(14):3476-3483
[78]Levison Peter R., Badger Stephen E., Dennis Jon, et al. Recent developments of magnetic beads for use in nucleic acid purification[J]. Journal of Chromatography A,1998, 816:107-111
[79]Levison Peter R., Badger Stephen E., Dennis Jon, et al. New approaches to the isolation of DNA by ion-exchange chromatography[J]. Journal of Chromatography A,1998,827: 337-344
[80]Brandon Yoza, Mitsufumi Matsumoto, Tadashi Matsunaga. DNA extraction using modified bacterial magnetic particles in the presence of amino silane compound[J]. jounal of Biotechnology,2001,94:217-224
[81]Brandon Yoza, Atsushi Arakaki, Tadashi Matsunaga. DNA extraction using bacterial magnetic particles modified with hyperbranched polyamidoamine dendrimer[J]. Journal of Biotechnology,2002,101:219-228
[82]孙敏莉,王强斌,张皓,et al. DEAE-磁粒纳米粒子提纯质粒的研究[J].上海第二医科大学学报,2003,23(6)：512-514
[83]Chen-Li Chiang, Ching-Shan Sung, Ting-Feng Wu, et al. Application of superparamagnetic nanoparticles in purification of plasmid DNA from bacterial cells[J]. Journal of Chromatography B,2005,822:54-60
[84]Matthew John Baker. Isolation of nucleic acids[P]. US patent:2007/0231892,2007-10-4