Size effects of magnetic beads in circulating tumour cells magnetic capture based on streptavidin–biotin complexation

Circulating tumour cells (CTCs) draw significant attention as a promising biomarker for cancer prognosis, status monitoring, and metastasis diagnosis. However, the concentration of CTCs in peripheral blood is usually extremely low, thereby requiring enrichment followed by isolation of CTCs prior to detection. An immunomagnetic separation is a promising tool for CTCs enrichment. In this study, a cost‐effective magnetic separation method, based on streptavidin–biotin complexation, was developed and the effects of magnetic beads’ size in CTCs capture were compared. Magnetic nanobeads which were 25 nm in diameter lead to highest capture efficiency (82.2%) compared with 150 nm magnetic beads and 1 µm microbeads. Based on the streptavidin–biotin system, 25 nm magnetic nanobeads could capture model CTCs over 80% efficiency even at concentrations as low as ∼25 cells/mL that may represent the actual level of CTCs in peripheral blood of cancer patients. Furthermore, the isolated cells remained robust and healthy showing insignificant changes in morphology and behaviour when cultured for 24 h immediately after capture and isolation. The magnetic nanobeads based on streptavidin–biotin complexation showed promise for the easy and efficient capture and isolation of healthy CTCs for further diagnosis and analysis.Inspec keywords: cancer, magnetic separation, nanomedicine, nanomagnetics, proteins, biomagnetism, tumours, cellular biophysics, magnetic particles, molecular biophysics, blood, nanoparticlesOther keywords: streptavidin–biotin complexation, cancer prognosis, peripheral blood, immunomagnetic separation, CTCs capture, streptavidin–biotin system, circulating tumour cells, CTC enrichment, magnetic separation method, magnetic nanobeads, magnetic capture, size 25.0 nm, size 150.0 nm, time 24.0 hour     

Study of flow fractionation characteristics of magnetic chromatography utilizing high-temperature superconducting bulk magnet

Abstract

We present numerical simulation of separating magnetic particles with different magnetic susceptibilities by magnetic chromatography using a high-temperature superconducting bulk magnet. The transient transport is numerically simulated for two kinds of particles having different magnetic susceptibilities. The time evolutions were calculated for the particle concentration in the narrow channel of the spiral arrangement placed in the magnetic field. The field is produced by the highly magnetized high-temperature superconducting bulk magnet. The numerical results show the flow velocity difference of the particle transport corresponding to the difference in the magnetic susceptibility, as well as the possible separation of paramagnetic particles of 20 nm diameter.     

Study of flow fractionation characteristics of magnetic chromatography utilizing high-temperature superconducting bulk magnet

We present numerical simulation of separating magnetic particles with different magnetic susceptibilities by magnetic chromatography using a high-temperature superconducting bulk magnet. The transient transport is numerically simulated for two kinds of particles having different magnetic susceptibilities. The time evolutions were calculated for the particle concentration in the narrow channel of the spiral arrangement placed in the magnetic field. The field is produced by the highly magnetized high-temperature superconducting bulk magnet. The numerical results show the flow velocity difference of the particle transport corresponding to the difference in the magnetic susceptibility, as well as the possible separation of paramagnetic particles of 20 nm diameter.     

Dynamic capture and accumulation of multiple types of magnetic particles based on fully coupled multiphysics model in multiwire matrix for high-gradient magnetic separation

High-gradient magnetic separation (HGMS) effectively separates fine weakly magnetic minerals using a magnetic matrix. The basic principle of single-wire capture of magnetic particles in HGMS has received considerable attention. In practice, however, a real matrix is made of numerous magnetic wires. Transport of magnetic particles inside a multiwire matrix under various operating conditions has not been sufficiently investigated, and it is not clear whether single-wire and multiwire matrices differ significantly. A fully coupled multiphysics model based on the idealized capture model was developed to investigate the 2D capture and accumulation of multiple types of particles in single-wire and multiwire matrices. In this model, the properties of multiple types of particles were defined. Then, particle tracing via the fluid flow model was used to calculate the dynamic capture and accumulation of particles under the determined magnetic and flow fields. The time-dependent dynamic capture mode used in this study can reveal the details of particle capture and accumulation in single-wire and multiwire matrices. All the calculations and analyses indicate that single-wire and multiwire matrices both exhibit basically the similar capture tendency as the particle size, slurry feed velocity, and magnetic induction are gradually increased, and a single-wire matrix always has a much higher capture selectivity than a multiwire matrix. This difference in selectivity between the single-wire and multiwire matrices results mainly from magnetic coupling between magnetic wires in the multiwire matrix, where the fluid flow is also quite complicated. In addition, adjacent columns of wires are staggered vertically, increasing the probability of collisions between the particles and the wires; thus, intergrowth particles that are not captured by the upstream wires are more easily captured by the downstream wires. By comparing the experimental results with the simulation results, the correctness of the HGMS recovery and grade prediction results was verified.     

Control of lattice spacing in a triangular lattice of feeble magnetic particles formed by induced magnetic dipole interactions

Abstract

We studied methods of controlling the spacing between particles in the triangular lattice formed by feeble magnetic particles through induced magnetic dipole interaction. Formation of a triangular lattice is described by the balance between the magnetic force and the interaction of induced magnetic dipoles. The intensity of the magnetic force is proportional to the volume of particles V and the difference in the magnetic susceptibilities between the particles and the surrounding medium Δχ. On the other hand, the intensity of the induced magnetic dipole interaction depends on the square of V and Δχ. Therefore, altering the magnetic susceptibility difference by changing the susceptibility of the surrounding medium, volume of the particles, and intensity and spatial distribution of the applied magnetic field effectively controls the distance between the particles. In this study, these three methods were evaluated through experiment and molecular dynamics simulations. The distance between the particles, i.e. the lattice constant of the triangular lattice, was varied from 1.7 to 4.0 in units of the particle diameter. Formation of self-organized triangular lattice through the induced magnetic dipole interaction is based on magnetism, a physical property that all materials have. Therefore, this phenomenon is applicable to any materials of any size. Consequently, structure formation through induced magnetic dipole interaction is a potential way of fabricating materials with ordered structures.     

Intermethod comparison of the particle size distributions of colloidal silica nanoparticles

There can be a large variation in the measured diameter of nanoparticles depending on which method is used. In this work, we have strived to accurately determine the mean particle diameter of 30–40 nm colloidal silica particles by using six different techniques. A quantitative agreement between the particle size distributions was obtained by scanning electron microscopy (SEM), and electrospray-scanning mobility particle sizer (ES-SMPS). However, transmission electron microscopy gave a distribution shifted to smaller sizes. After confirming that the magnification calibration was consistent, this was attributed to sample preparation artifacts. The hydrodynamic diameter, d _h , was determined by dynamic light scattering (DLS) both in batch mode, and hyphenated with sedimentation field flow fractionation. Surprisingly the d _h were smaller than the SEM, and ES-SMPS diameters. A plausible explanation for the smaller sizes found with DLS is that a permeable gel layer forms on the particle surface. Results from nanoparticle tracking analysis were strongly biased towards larger diameters, most likely because the silica particles provide low refractive index contrast. Calculations confirmed that the sensitivity is, depending on the shape of the laser beam, strongly size dependent for particles with diameters close to the visualization limit.     

An extracellular polymer at the interface of magnetic bioseparations

FucoPol, a fucose-containing extracellular polysaccharide (EPS) produced by bacterium Enterobacter A47 using glycerol as the carbon source, was employed as a coating material for magnetic particles (MPs), which were subsequently functionalized with an artificial ligand for the capture of antibodies. The performance of the modified MPs (MP–EPS-22/8) for antibody purification was investigated using direct magnetic separation alone or combined with an aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) and dextran. In direct magnetic capturing, and using pure protein solutions of human immunoglobulin G (hIgG) and bovine serum albumin (BSA), MP–EPS-22/8 bound 120 mg hIgG g⁻¹ MPs, whereas with BSA only 10 ± 2 mg BSA g⁻¹ MPs was achieved. The hybrid process combining both the ATPS and magnetic capturing leads to a good performance for partitioning of hIgG in the desired phase as well as recovery by the magnetic separator. The MPs were able to bind 145 mg of hIgG g⁻¹ of particles which is quite high when compared with direct magnetic separation. The theoretical maximum capacity was calculated to be 410 ± 15 mg hIgG adsorbed g⁻¹ MPs with a binding affinity constant of 4.3 × 10⁴ M⁻¹. In multiple extraction steps, the MPs bound 92% of loaded hIgG with a final purity level of 98.5%. The MPs could easily be regenerated, recycled and re-used for five cycles with only minor loss of capacity. FucoPol coating allowed both electrostatic and hydrophobic interactions with the antibody contributing to enhance the specificity for the targeted products.     

Synthesis and magnetic properties of Ni–SiO2 nanocomposites

Nanocomposite Ni_(1 − x)/(SiO₂)_x soft magnetic materials were synthesized by a simple sol–gel combined hydrogen reduction method. The crystal structure of the particles was determined by X-ray diffraction (XRD). The shapes and sizes of the metal particles embedded in the SiO₂ matrix were determined by transmission electron microscopy (TEM), and magnetic properties were measured by the vibrating samples magnetometer (VSM). The obtained nanocomposite material is composed of nanoparticles coated with a thin SiO₂ layer, and with the content of the silicon increase, the thickness of the silica shells increase and the saturation magnetization decrease. The diameter of Ni particle in the sample is about 30–40 nm. The influence of the Ni content and preparation conditions on the microstructures and magnetic properties were discussed.     

Quantitative Magnetic Separation of Particles and Cells Using Gradient Magnetic Ratcheting

Extraction of rare target cells from biosamples is enabling for life science research. Traditional rare cell separation techniques, such as magnetic activated cell sorting, are robust but perform coarse, qualitative separations based on surface antigen expression. A quantitative magnetic separation technology is reported using high‐force magnetic ratcheting over arrays of magnetically soft micropillars with gradient spacing, and the system is used to separate and concentrate magnetic beads based on iron oxide content (IOC) and cells based on surface expression. The system consists of a microchip of permalloy micropillar arrays with increasing lateral pitch and a mechatronic device to generate a cycling magnetic field. Particles with higher IOC separate and equilibrate along the miropillar array at larger pitches. A semi‐analytical model is developed that predicts behavior for particles and cells. Using the system, LNCaP cells are separated based on the bound quantity of 1 μm anti‐epithelial cell adhesion molecule (EpCAM) particles as a metric for expression. The ratcheting cytometry system is able to resolve a ±13 bound particle differential, successfully distinguishing LNCaP from PC3 populations based on EpCAM expression, correlating with flow cytometry analysis. As a proof‐of‐concept, EpCAM‐labeled cells from patient blood are isolated with 74% purity, demonstrating potential toward a quantitative magnetic separation instrument.     

TECHNIQUES FOR SELECTIVE MAGNETIC SEPARATION OF FINE PARTICLES

ABSTRACT

Magnetic separation as a particle-particle or particle-fluid separation technique has been extended to be effective for particulates with the smallest known magnetic susceptibility. Most commonly found materials are diamagnetic but the effectiveness of high magnetic field gradient separators for even these very weakly magnetic materials makes it difficult to separate more magnetic species of particles from them. The selectivity of such separations has been improved by matrix design and by several separation techniques. Regular arrays of matrix wires can be arranged according to the calculated field profile to exclude regions of capture for magnetic particulates of positive or negative susceptibility. The magnetic field orientation with respect to the array provides control over the competition between magnetic capture forces and those of fluid flow. The size of particle depletion regions in model arrays depends on particle size and susceptibility and suggests a method of measurement of these even for submicron particulates.     

A novel pH-sensitive magnetic alginate–chitosan beads for albendazole delivery

Background: Drug delivery system using polymer-coated magnetic carriers is considered as an effective strategy for passive targeting, which can not only increase drug utilization but also reduce the adverse reaction. With the carriers, sensitivity to physical stimuli (e.g., magnetic field, pH) has been developed and drugs were conjugated to form incorporating magnetic particles, so that drugs could be located to desire position. Method: Novel magnetic alginate (Alg)–chitosan (CS) beads loaded with albendazole (ABZ) were prepared and evaluated for pH sensitivity and drug release characteristics. The effects of six different factors (Alg concentration, the weight ratio of drug to polymer, the weight ratio of magnetite nanoparticles to polymer, CaCl2 concentration, CS concentration, the volume ratio of Alg to CS) were studied on the swelling ability of the magnetic beads. The magnetic beads were characterized by Fourier transform infrared spectroscopy, scanning electron microscope, and vibrating sample magnetometry. In addition, the delivery behavior of ABZ from the magnetic beads was studied. Result: The magnetic Alg–CS beads had showed unique pH-dependent swelling behaviors and a continuous release of ABZ. From the magnetometer measurements data, the beads also had superparamagnetic property as well as fast magnetic response. Conclusion: The pH-sensitive magnetic beads may be used as a magnetic drug targeting system for ABZ in the gastrointestinal tract.     

Efficient Capture and Simple Quantification of Circulating Tumor Cells Using Quantum Dots and Magnetic Beads

Circulating tumor cells (CTCs) are valuable biomarkers for monitoring the status of cancer patients and drug efficacy. However, the number of CTCs in the blood is extremely low, and the isolation and detection of CTCs with high efficiency and sensitivity remain a challenge. Here, we present an approach to the efficient capturing and simple quantification of CTCs using quantum dots and magnetic beads. Anti‐EpCAM antibody‐conjugated quantum dots are used for the targeting and quantification of CTCs, and quantum‐dot‐attached CTCs are isolated using anti‐IgG‐modified magnetic beads. Our approach is shown to result in a capture efficiency of about 70%–80%, enabling the simple quantification of captured CTCs based on the fluorescence intensity of the quantum dots. The present method can be used effectively in the capturing and simple quantification of CTCs with high efficiency for cancer diagnosis and monitoring.     

SUBMICRON MAGNETIC PARTICLE SEPARATION

ABSTRACT

The control, collection or separation of particles on the basis of Their magnetic moment relative Co the carrier fluid has been demonstrated in many applications. Usually the particle sizes are larger than one micron and the magnetic susceptibility at least moderately paramagnetic. Recently, particle separation techniques have been developed for both diamagnetic and submicron particles. These techniques have found application in mineral beneficiation, nuclear reactor coolants, biology and medicine. Such developments require an understanding of flow forces in liquids and gases, diffusion and Brownian motion, and of magnetic properties which range from the strong magnetic moments of ferromagnetic and superparamagnetic particles down orders of magnitude to those of diamagnetism.     

Effect of the Magnetic Dipole–Dipole Interaction on the Capture Efficiency in Open Gradient Magnetic Separation

In magnetic separation, the magnetic dipole-dipole (DD) interaction between particles has an important effect on the capture efficiency. By producing transient particle agglomerations, this interaction can considerably speed up the separation process. To take into account adequately this effect in ferromagnetic particle random dispersion, we have developed a modeling approach. The approach is based on the coupling of the magnetic force equation and a local homogenizing model for the material magnetic permeability. To verify the efficiency of the proposed approach on one hand and to estimate the effect of the DD interaction on the particle capture on the other hand, we consider a problem of open gradient magnetic separation (OGMS). We also conducted a limited experimental verification of the transient agglomeration for fine ferromagnetic particles.     

Mathematical modeling of coating time in dry particulate coating using mild vibration field with bead media described by DEM simulation

To theoretically understand the previously reported dry particulate coating process using a mild vibration field with a bead media, a mathematical analysis model of the dry coating system was developed. In this coating process, an ordered mixture with coarse host particles (drug-loaded ion exchange resin, diameter approximately 100 µm) and fine guest particles (acrylic polymer particle, primary particle size of approximately 100 nm) is formed using a vibrating a vessel. Second, the guest particles on the host particulate surface are firmly fixed using the collision of coated particles zirconia beads (diameter 1.5 mm). Our model assumes that the unfixed guest particles are fixed by particle-to-particle collisions (C_c) provided by the apparatus, thereby increasing the coating ratio. C_c was estimated using the discrete element method and some experimental results. The model includes parameters such as the number of C_c, host particles and unfixed guest particles. The coating time simulated by the established model equation in this study fits well with the experimental results of the dry process. It depends on the ratio of the number of collisions contributing to the increased coating ratio to the number of unfixed guest particles on the surface of host particles.     

On the motion of superparamagnetic particles in magnetic drug targeting

A stochastic ODE model is developed for the motion of a superparamagnetic cluster suspended in a Hagen-Poiseuille flow and guided by an external magnet to travel to a target. The specific application is magnetic drug targeting, with clusters in the range of 10–200 nm radii. As a first approximation, we use a magnetic dipole model for the external magnet and focus on a venule of 10⁻⁴ m radius close to the surface of the skin as the pathway for the clusters. The time of arrival at the target is calculated numerically. Variations in release position, background flow, magnetic field strength, number of clusters, and stochastic effects are assessed. The capture rate is found to depend weakly on variations in the velocity profile, and strongly on the cluster size, the magnetic moment, and the distance between the magnet and the blood vessel wall. A useful condition is derived for the optimal capture rate. The case of simultaneous release of many clusters is investigated. Their accumulation in a neighborhood of the target at the venule wall follows a normal distribution with the standard deviation roughly half of the distance between the magnet and the target. Ideally, this deviation should equal the tumor radius, and the magnet should be beneath the center of the tumor. The optimal injection site for a cluster is found to be just prior to arrival at the target. Two separate mechanisms for capturing a cluster are the magnetic force and, for radii smaller than 20 nm, Brownian motion. For the latter case, the capture rate is enhanced by Brownian motion when the cluster is released near the wall.     

Nanocrystalline magnetic alloys and ceramics

Magnetic properties of materials in their nanocrystalline state have assumed significance in recent years because of their potential applications. A number of techniques have been used to prepare nanocrystalline magnetic phases. Melt spinning, high energy ball milling, sputtering, glassceramization and molecular beam epitaxy are some of the physical methods used so far. Among the chemical methods, sol-gel and co-precipitation routes have been found to be convenient. Ultrafine particles of both ferro- and ferrimagnetic systems show superparamagnetic behaviour at room temperature. Coercivity(H _c ) and maximum energy product(BH) _max of the magnetic particles can be changed by controlling their sizes. The present paper reviews all these aspects in the case of nanocrystalline magnetic systems — both metallic and ceramics.     

Concentration of Bacillus spores by using silica magnetic particles

Silica particles are mainly used for the concentration of nucleic acid for diagnostic purposes. This is usually done under acidic or chaotropic conditions that will demolish most of the living organisms and prevent the application of other diagnostic tests. Here we describe the development of a method for the capturing and concentration of Bacillus spores using silica magnetic particles to enable fast and sensitive detection. We have shown that capturing various Bacilli spores via silica magnetic particles is limited, with large differences between spore batches (42 +/- 25%). The hydrophobic exosporium layer of spore limits the capture by the hydrophilic silica beads. Partial removal of Bacillus exosporium increases capture efficiency. To increase capturing efficiency without harming the spores' viability, a cationic lipid, didecyldimethylammonium bromide (DDAB), was used as a coat for the negatively charged silica particles. DDAB treatment increased capture efficiency from 42% to more than 90%. Using this method, we were able to capture as few as 100 Bacillus anthracis spores/mL with 90% efficacy. Release of captured spores was achieved by the addition of albumin. The capture and release processes were verified by plating and by flow cytometry using light scatter analysis. The method is simple, efficient, easy to operate, and fast.     

Influence of electrophoresis waveforms in determining stochastic nanoparticle capture rates and detection sensitivity

Electrophoretic capture and release of charged polystyrene particles at the opening of a membrane pore has been investigated to determine the optimal applied current waveform, iapp(t), for ensuring true stochastic counting rates and to improve detection sensitivity (i.e., Delta(counts per second)/Delta(particle concentration)). In capture and release detection, charged particles are electrophoretically driven to the opening of a small pore ( approximately 60 nm diameter) in a membrane; capture of a single particle at the pore opening at time tau is signaled by a decrease in the flux of a redox species (Fe(CN)(6)4-) through the pore. The captured particle is then released by applying an electrophoretic current in the opposite direction, and the process is repeated to acquire sufficient statistical data to determine the solution particle concentration (Cp) based on the relationship between Cp and the average particle counting rate (tauavg(-1)). Both tauavg(-1) and the method sensitivity are shown, for the detection of 90 nm diameter polystyrene particles, to depend strongly on the applied current waveform. The observed dependence is a consequence of the nonequilibrium distribution of particles in the analyte solution that results from electrostatic forces acting on the particle whenever iapp has a nonzero value. Stochastic capture rates corresponding to an initial uniform distribution of particles are more closely achieved using an applied current waveform that includes an equilibration period (iapp=0) prior to electrophoretic capture. An increase in particle detection sensitivity, relative to the previously reported value, results from this equilibration step.     

Modelling of filtration processes of fibrous filter media

This work presents a stochastic approach, based on Monte Carlo method, to simulate liquid filtration processes through non-woven fibrous materials. The real filter material is represented as a multilayer medium with a network of multiply connected pores. To describe the deposition and resuspension of particles on and from the filter medium, the following four mechanisms were considered: particle capture by sieving, patricle capture by fibers; particle capture by blocked pores; and particle re-entrainment. The particle capture by fibers and blocked pores, and particle re-entrainment depend on the balance between the adhesion and removal forces. The adhesion forces for particles of diameter smaller than 20 μm were determined through the concept of London-Van Der Waals forces. For particles of diameter greater than 20 μm, gravitational forces were considered. Three-dimensional random flow was assumed to stimulate the particles motion through the multilayer medium. The pressure drop across the filter medium was calculated as the sum of the pressure drop across the clean filter plus the pressure drop due to the deposited particles.A FORTRAN Program was developed to implement the filtration process model. For a wide range of typical filtration conditions, the calculated filter efficiencies predicted the experimental results with a percent difference between 0.5 and 19.3 depending on the particle size. The filter material capacities were predicted with an average discrepancy of 23.0%