磁性纳米颗粒在生物医学领域中的应用

磁性纳米颗粒作为一种新型纳米材料,在许多领域,特别是在生物医药、生物工程等方面具有广阔的应用前景.本文着重论述了近年来磁性纳米颗粒在生物分离、靶向给药、热疗以及磁共振成像对比剂等方面的应用,并对其应用前景进行了展望.     

Different methods for the fractionation of magnetic fluids

 Magnetic fluids are used in many fields of application, such as material separation and biomedicine. Magnetic fluids consist of magnetic nanoparticles, which commonly display a broad distribution of magnetic and nonmagnetic parameters. Therefore, upon application only a small number of particles contribute to the desired magnetic effect. In order to optimize magnetic fluids for applications preference is given to methods that separate magnetic nanoparticles according to their magnetic properties. Hence, a magnetic method was developed for the fractionation of magnetic fluids. Familiar size-exclusion chromatography of two different magnetic fluids was carried out for comparison. The fractions obtained and the original samples were also magnetically characterized by magnetic resonance and magnetorelaxometry, two biomedical applications. The size-exclusion fractions are similar to those of magnetic fractionation, despite the different separation mechanisms. In this respect, magnetic fractionation has several advantages in practical use over size-exclusion chromatography: the magnetic method is faster and has a higher capacity. The fractions obtained by both methods show distinctly different magnetic properties compared to the original samples and are therefore especially suited for applications such as magnetorelaxometry. Received: 12 July 1999/Accepted in revised form: 9 November 1999     

Magnetoanalysis of micro/nanoparticles: a review

Application of magnetic field on the separation and analysis of nano/microparticles is a growing subject in analytical separation chemistry. The migration phenomenon of a particle under inhomogeneous magnetic field is called magnetophoresis. The migration velocity depends on the magnetic susceptibility and the size of a particle. Therefore, magnetophoresis allows us to determine the magnetic susceptibility of particles, and to separate particles based on the magnetic properties. Magnetic separation of ferromagnetic particles in liquid has been utilized for a long time. For example, a high gradient magnetic separation under the non-uniform magnetic field generated by ferromagnetic mesh has been utilized in a wide region from chemical industry to bioscience. Recent progress on magnetic nanoparticles and microfluidic devices has made it possible to extend the range of application. Furthermore, it has been demonstrated that the very sensitive measurement of the magnetic susceptibility of microparticles can be performed by observing magnetophoretic velocity. In this review, we mainly introduce novel separation and detection methods based on magnetophoresis, which have been invented in this decade, and then new principles of particle migration under magnetic field are presented.     

磁性微球及其在免疫学中应用的研究进展

磁性微球作为一种新型功能高分子材料,在生物医学领域有着广泛的应用前景。特别是在免疫学方面,常用于细胞分离、疾病诊断、食品检测等,并取得了显著的科研成果。本文介绍了磁性微球常见的制备和表面改性方法,并着重综述了其在免疫学中应用的研究进展。     

磁性微珠的制备及其在生物样品分离富集中的应用

磁性微珠作为一种新型的功能材料,广泛应用于磁性材料、生物医学、细胞学、生物工程及分离工程等诸多领域,并显示出强大的生命力。本文对磁性微珠的结构、性质、制备方法及其在生物样品的分离富集等方面的应用,进行了总结和评述,引用文献62篇。     

Field-flow fractionation of magnetic particles in a cyclic magnetic field

Although magnetic field-flow fractionation (MgFFF) is emerging as a promising technique for characterizing magnetic particles, it still suffers from limitations such as low separation efficiency due to irreversible adsorption of magnetic particles on separation channel. Here we report a novel approach based on the use of a cyclic magnetic field to overcome the particle entrapment in MgFFF. This cyclic field is generated by rotating a magnet on the top of the spiral separation channel so that magnetic and opposing gravitational forces alternately act on the magnetic particles suspended in the fluid flow. As a result, the particles migrate transversely between the channel walls and their adsorption at internal channel surface is prevented due to short residence time which is controlled by the rotation frequency. With recycling of the catch-release process, the particles follow saw-tooth-like downstream migration trajectories and exit the separation channel at velocities corresponding to their sedimentation coefficients. A retention model has been developed on the basis of the combined effects of magnetic, gravitational fields and hydrodynamic flow on particle migration. Two types of core-shell structured magnetic microspheres with diameters of 6.04- and 9.40-μm were synthesized and used as standard particles to test the proposed retention theory under varying conditions. The retention ratios of these two types of particles were measured as a function of magnet rotation frequency, the gap between the magnet and separation channel, carrier flow rate, and sample loading. The data obtained confirm that optimum separation of magnetic particles with improved separation efficiency can be achieved by tuning rotation frequency, magnetic field gradient, and carrier flow rate. In view of the widespread applications of magnetic microspheres in separation of biological molecules, virus, and cells, this new method might be extended to separate magnetically labeled proteins or organisms for multiplex analyte identification and purification.     

基于识别分子的磁分离技术在食源性致病菌分离中应用

食源性致病菌是一类对人类健康危害极大的致病菌,但由于其在食品基质中数量较少,通常需要在检测前对其进行富集。磁分离作为一种常用的前处理手段,在致病菌的富集、分离方面展示出极好的应用前景。常用于修饰在磁性材料上的识别分子有多种,本文主要综述了抗体、抗生素、核酸、凝集素等主要的几种识别分子在磁分离中的应用。     

高性能磁性纳米粒子的有机相制备与生物应用的研究进展

磁性纳米粒子(MNPs)的合成开发在基础科学研究和技术应用方面得到了深入的发展。与大块的磁性材料不同,MNPs展现出了独特的磁性,并且可以通过系统的纳米尺寸工程调控它们的性能。本文首先简要介绍了MNPs的基本特征,总结了不同MNPs的制备方法,包括金属、合金、金属氧化物和多功能的MNPs;重点关注了可精确控制MNPs尺寸、形状、组成和结构的有机相合成方法;最后讨论了这些MNPs在生物方面的应用。     

Review: Preparation and Application of Magnetic Chitosan Derivatives in Separation Processes

Chitosan is one of the most abundant natural polysaccharide in nature. Due to its unique properties, chitosan has fascinated the scientific community since its discovery. When modified with other materials and combined with magnetic particles, the resulting composite material, a magnetic chitosan derivative, is provided with three significant characteristics. First, chitosan has excellent properties for preconcentration/extraction, such as adsorption and chelating effects, low cost, and nontoxicity. Second, new functional groups have enhanced the properties of chitosan that include water solubility, stability, recyclability, and enhanced adsorption capacity. Finally, due to the efficient and fast adsorption processes, as well as simple and convenient magnetic separation, the magnetic adsorbents greatly reduce the time of sample handling. In this article, recent synthesis and modification methods of magnetic chitosan derivatives are reviewed along with some applications in analytical separations.     

Facile synthesis and application of teicoplanin‐modified magnetic microparticles for enantioseparation

A facile, mild method for the preparation of teicoplanin (TE) modified Fe₃O₄ microparticles (MPs) employing polydopamine (PDA) layer as a versatile secondary reaction platform was developed. The synthesized magnetic MPs (Fe₃O₄@PDA@TE) were characterized by various characterization techniques, such as TEM, zeta potential etc, to affirm the successful modification of TE to magnetic Fe₃O₄ MPs. Using the as‐synthesized materials as chiral adsorbents, efficient chiral separation of representative racemic compounds was successfully achieved. Due to the magnetic responsivity, the materials were easily isolated from the racemic solutions under an external magnetic field and could be readily reused for at least three times. Thus, the well‐prepared functional magnetic MPs have great potential in preparative chiral separation.     

基于侧向磁泳的微流控芯片对两种磁性纳米球的同时分选

基于磁性纳米球在微流控芯片上的侧向磁泳,利用微流控芯片分选了不同磁响应性的磁球.提出了包含磁性纳米球聚集与偏移的理论模型,用于分析磁球在芯片上的侧向位移.在理论分析的基础上设计了芯片系统,使不同磁响应性的磁纳米球可以在芯片系统上依次被分选.实验结果表明,2种磁性纳米球的分选效率均可接受,且实验操作简单;磁响应性强的磁球可被完全分离,这对于珍贵分析样品的分选很有价值.该分选系统被成功用于同时分选样品中乙型肝炎病毒的DNA与丙型肝炎病毒的反转录DNA,在生化分析中具有广阔的应用前景.     

基于贻贝仿生化学的分离功能材料

贻贝仿生的表面化学是近年来材料学、化学、生物医学等领域的交叉研究热点。多巴胺可以作为贻贝足丝蛋白(Mfp)超强黏附特性的模型分子,通过复杂的氧化-自聚和组装,形成多种功能的聚多巴胺(PDA)纳米涂层和纳米粒子,在分离膜、吸附材料、生物医用材料、生物黏结剂等领域有着广阔的应用前景。本研究小组近年来持续开展了基于贻贝仿生化学的分离功能材料制备与结构调控的研究工作,率先将多巴胺表面沉积方法应用于多孔分离膜表面的构建与功能化,提出了多巴胺的自聚-沉积过程模型,进而验证了PDA沉积层的纳滤分离特性,建立了一条简单方便的膜表面功能化与纳滤膜制备新途径。本文主要对基于贻贝仿生化学的分离功能材料,特别是分离膜的研究进展进行综述,并对将来的发展趋势进行展望。     

Integration of carboxyl modified magnetic particles and aqueous two-phase extraction for selective separation of proteins

Both of the magnetic particle adsorption and aqueous two-phase extraction (ATPE) were simple, fast and low-cost method for protein separation. Selective proteins adsorption by carboxyl modified magnetic particles was investigated according to protein isoelectric point, solution pH and ionic strength. Aqueous two-phase system of PEG/sulphate exhibited selective separation and extraction for proteins before and after magnetic adsorption. The two combination ways, magnetic adsorption followed by ATPE and ATPE followed by magnetic adsorption, for the separation of proteins mixture of lysozyme, bovine serum albumin, trypsin, cytochrome C and myloglobin were discussed and compared. The way of magnetic adsorption followed by ATPE was also applied to human serum separation.     

Microchip electrophoresis coupled with on-line magnetic separation and chemiluminescence detection for multiplexed immunoassay

A facile and universal strategy for multiplexed immunoassay is proposed. The strategy is based on microchip electrophoresis (MCE) coupled with on-line magnetic separation and chemiluminescence (CL) detection. The system consisted of a microchip, an electromagnet, and a photomultiplier. The realization of multiplexed immunoassay protocol involves sampling magnetic nanoparticles (MNPs) labeled antibodies, N-(4-aminobutyl)-N-ethyl-isoluminol (ABEI) labeled antigens and free antigens in the precolumn reactor, on-line immunoreaction, capturing the MNPs-immunocomplexes, and the separation of unconjugated ABEI-labeled antigens. After on-line magnetic separation, the free ABEI-labeled antigens were transported into the separation channel, and mixed with hydrogen peroxide (H(2) O(2) ) in the presence of horseradish peroxidase in the postcolumn reactor, and producing CL emission. Using this arrangement, multiple analytes could be measured simultaneously by performing the technical operations for a single assay. As a proof-of-concept, the multiplexed immunoassay was evaluated for the simultaneous determination of five model analytes (i.e. hydrocortisone, corticosterone, digoxin, testosterone, and estriol). The results exhibited excellent precision and sensitivity, the relative standard deviations for nine times detection were lower than 4.7% for all the five components, and the detection limits of five analytes were in the range of 3.6-4.9 nM. The MCE system was validated using two human serum-based control samples containing five analytes.     

Optimization of adsorption conditions of BSA on thermosensitive magnetic composite particles using response surface methodology

Thermosensitive core-shell magnetic composite particles with a magnetic silica core and a rich poly (N-vinylcaprolactam) (PNVCL) shell layer were developed for studying the adsorption of bovine serum albumin (BSA) in a batch system. Various analytical and spectroscopic techniques including SEM, FT-IR, VSM and DSC were used to characterize the adsorbents prepared in this study. The combined effects of operating parameters such as initial temperature, pH and initial BSA concentration on the adsorption were analyzed using response surface methodology. The optimum conditions were 40°C, pH 4.68, and initial BSA concentration 2.0 mg/mL. Desorption experiments were conducted by altering the system temperature where a high recovery rate of protein was obtained. The separation process developed here indicates that the dual-responsive smart adsorbent could be an ideal candidate for the separation of protein.     

Affinity separation of enzymes from mixtures containing suspended solids

Agarose beads containing immobilized enzymes or affinity ligands have been made magnetically responsive by adsorbing freshly precipitated magnetite on their surface. These beads are used for affinity adsorption of proteins from complex mixtures containing suspended solids. The magnetically responsive beads and the unwanted (diamagnetic) solids are then separated by magnetic filtration. This magnetic adsorption scheme for direct affinity separation of enzymes from mixtures containing suspended solids is compared with a similar, but nonmagnetic, scheme in which the affinity matrix is supported on fiberglass cloth. The enzyme is allowed to adsorb in this matrix, and the matrix is simply removed physically from the suspension to achieve separation from the unwanted solids. The two methods seem comparable in their ability to separate a desired enzymatic activity. The magnetic methods are technically the more complex of the two, but are significantly the more rapid. The efficiency of separation of diamagnetic and ferrimagnetic solids in these biological systems by high gradient magnetic filtration is good.     

Synthesis and Applications of Functionalized Magnetic Nanomaterials in Enantioseparation

Efficient enantioselective separation is a challenging task due to the identical physical and chemical properties of enantiomers. Functionalized magnetic nanomaterials modified with chiral ligands on their surface possess both magnetic property and chiral recognition ability, and have demonstrated great potential in chiral discrimination. This review summarizes the applications of magnetic nanomaterials modified with various chiral selectors (e.g., β-cyclodextrins, polymers, proteins, amino acids and cellulose) in enantioselective separation. After proper preparation and modification, these functionalized magnetic nanomaterials are effective for enantioseparation. Therefore, enantioseparations based on functionalized magnetic nanomaterials are convenient, economical and effective.     

Superparamagnetic lysozyme surface-imprinted polymer prepared by atom transfer radical polymerization and its application for protein separation

Molecular imprinting as a promising and facile separation technique has received much attention because of their high selectivity for target molecules. In this study, the superparamagnetic lysozyme surface-imprinted polymer was prepared by a novel fabricating protocol, the grafting of the imprinted polymer on magnetic particles in aqueous media was done by atom transfer radical polymerization (ATRP), and the properties of the imprinted polymer were characterized in detail. Its high selective adsorption and recognition to lysozyme demonstrated the separation ability of the magnetic imprinted material to template molecule, and it has been used for quick and direct separation of lysozyme from the mixture of standard proteins and real egg white samples under an external magnetic field. Furthermore, the elution of lysozyme from the imprinted material was achieved by PEG/sulphate aqueous two-phase system, which caused lysozyme not only desorption from the imprinted materials but also redistribution in the top and bottom phase of aqueous two-phase system. The aqueous two-phase system exhibited some of the extraction and enrichment effect to desorbed lysozyme. Our results showed that ATRP is a promising method for the protein molecularly imprinted polymer preparation.     

Preparation of magnetic hydroxyapatite clusters and their application in the enrichment of phosphopeptides

A novel strategy for the effective enrichment of phosphopeptides based on magnetic hydro‐xyapatite (HAp) clusters was developed in the current study. The structure of HAp ensures its probable separation capability, including cation exchange with P‐sites (negatively charged pairs of crystal phosphates), calcium coordination, anion exchange with C‐sites (positively charged pairs of crystal calcium ions). The prepared magnetic HAp clusters showed good performance on the efficient enrichment of phosphopeptides from the digestion mixture of β‐casein and BSA. Compared to commercial HAp particles, the magnetic HAp clusters exhibited better selectivity toward phosphopeptides. In addition, the use of magnetic material greatly simplified the enrichment procedure, which avoided the tedious centrifugation steps in a typical phosphopeptides enrichment protocol. Finally, the material was successfully applied in the enrichment of phosphopeptides from human serum. Taken together, the efficient enrichment of the phosphopeptides by the easily prepared magnetic HAp clusters demonstrated a rapid and convenient strategy for the purification of phosphopeptides from complex samples, which may facilitate protein phosphorylation studies.     

基于核酸适配体的新型荧光纳米生物传感器用于凝血酶的测定

利用凝血酶的两条核酸适配体与凝血酶的高亲和力构建了三明治结构, 利用磁性纳米颗粒的磁性分离技术, 设计并制作了一种新型的荧光纳米生物传感器, 用其检测凝血酶. 此法对凝血酶的响应线性范围为2.24×10-11～4.03×10-9 mol/L, 其线性方程为I=0.9758×1011c-2.628, 检出限为1.0×10-11 mol/L, 对浓度为2.68×10-10 mol/L的凝血酶检测10次, 其RSD为2.56％, 测得的荧光信号稳定, 24 h后测定并无衰减, 具有很高的检测特异性和灵敏度.