Cell affinity separations using magnetically stabilized fluidized beds: Erythrocyte subpopulation fractionation utilizing a lectin‐magnetite support

Putnam, David D.; Namasivayam, Vijay; Burns, Mark A.

doi:10.1002/bit.10511

Cited by 33 publications

(32 citation statements)

References 23 publications

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“…4 along with intersecting planes that represent sample volumetric flow rates utilized with magnet-activated and nonmagnetic cell separation systems described in the literature. Other researchers who have isolated cells using commercial 9 or microfluidic 5,22,25,[51][52][53][54] systems have employed volumetric flow rates on the order of 6.3-100 l min −1 . Figure 5 illustrates that this simple design can effectively meet and exceed the processing speeds or throughputs of both commercial systems ͑green plane͒ and microfluidic devices ͑red planes͒.…”

Section: B Microfluidic Device Design Optimizationmentioning

confidence: 99%

Computational design optimization for microfluidic magnetophoresis

2011

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Current macro-and microfluidic approaches for the isolation of mammalian cells are limited in both efficiency and purity. In order to design a robust platform for the enumeration of a target cell population, high collection efficiencies are required. Additionally, the ability to isolate pure populations with minimal biological perturbation and efficient off-chip recovery will enable subcellular analyses of these cells for applications in personalized medicine. Here, a rational design approach for a simple and efficient device that isolates target cell populations via magnetic tagging is presented. In this work, two magnetophoretic microfluidic device designs are described, with optimized dimensions and operating conditions determined from a force balance equation that considers two dominant and opposing driving forces exerted on a magnetic-particle-tagged cell, namely, magnetic and viscous drag. Quantitative design criteria for an electromagnetic field displacement-based approach are presented, wherein target cells labeled with commercial magnetic microparticles flowing in a central sample stream are shifted laterally into a collection stream. Furthermore, the final device design is constrained to fit on standard rectangular glass coverslip ͑60 ͑L͒ ϫ 24 ͑W͒ ϫ 0.15 ͑H͒ mm 3 ͒ to accommodate small sample volume and point-of-care design considerations. The anticipated performance of the device is examined via a parametric analysis of several key variables within the model. It is observed that minimal currents ͑Ͻ500 mA͒ are required to generate magnetic fields sufficient to separate cells from the sample streams flowing at rate as high as 7 ml/h, comparable to the performance of current state-of-the-art magnet-activated cell sorting systems currently used in clinical settings. Experimental validation of the presented model illustrates that a device designed according to the derived rational optimization can effectively isolate ͑ϳ100%͒ a magnetic-particle-tagged cell population from a homogeneous suspension even in a low abundance. Overall, this design analysis provides a rational basis to select the operating conditions, including chamber and wire geometry, flow rates, and applied currents, for a magnetic-microfluidic cell separation device.

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Section: B Microfluidic Device Design Optimizationmentioning

confidence: 99%

Computational design optimization for microfluidic magnetophoresis

2011

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“…Yields and purity can be comparable to FACS and MACS with reasonable throughput (10 8 -10 9 cells/h) (Putnam et al, 2003). To increase active surface area per unit volume, most macroscopic CAC systems adopt a packed bed design, giving rise to residence times in the order of 1-2 h (Putnam et al, 2003;Ujam et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

“…By immobilizing the antibodies directly onto surfaces, cell affinity chromatography (CAC) systems removes the need for pre-incubation while conserving the resolution and specificity of separation (Mandrusov et al, 1995;Putnam et al, 2003). Yields and purity can be comparable to FACS and MACS with reasonable throughput (10 8 -10 9 cells/h) (Putnam et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

“…Cell mixtures are fluorescently labeled using specific dyes or antibodies to surface antigens, and then sorted one by one based on light scattering and fluorescent properties. This technique results in highly pure cell populations that can be sorted by multiple parameters, at the expense of costly equipment and limited throughput ($10 7 cells/h) (Putnam et al, 2003;Thiel et al, 1998). Magnetic-activated cell sorting (MACS) is also used in the isolation of cells by a single parameter using antibody-coated magnetic beads.…”

Section: Introductionmentioning

confidence: 99%

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Enrichment using antibody‐coated microfluidic chambers in shear flow: Model mixtures of human lymphocytes

Sin

Murthy

Revzin

et al. 2005

Biotech & Bioengineering

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Isolation of phenotypically-pure cell subpopulations from heterogeneous cell mixtures such as blood is a difficult yet fundamentally important task. Current techniques such as fluorescent activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) require pre-incubation with antibodies which lead to processing times of at least 15-60 min. In this study, we explored the use of antibody-coated microfluidic chambers to negative deplete undesired cell types, thus obtaining an enriched cell subpopulation at the outlet. We used human lymphocyte cell lines, MOLT-3 and Raji, as a model system to examine the dynamic cell binding behavior on antibody coated surfaces under shear flow. Shear stress ranging between 0.75 and 1.0 dyn/cm2 was found to provide most efficient separation. Cell adhesion was shown to follow pseudo-first order kinetics, and an anti-CD19 coated (Raji-depletion) device with approximately 2.6 min residence time was demonstrated to produce 100% pure MOLT-3 cells from 50-50 MOLT-3/Raji mixture. We have developed a mathematical model of the separation device based on the experimentally determined kinetic parameters that can be extended to design future separation modules for other cell mixtures. We conclude that we can design microfluidic devices that exploits the kinetics of dynamic cell adhesion to antibody coated surfaces to provide enriched cell subpopulations within minutes of total processing time.

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“…Although single cell analysis is important, enriching rare cancer cells from a background of healthy cells must also be addressed. Among the many approaches to this problem, cell-affinity chromatography is one successful method for enriching rare cells [105][106][107]. In cell-affinity chromatography, a highaffinity ligand that binds selectively to a particular cell type is immobilized on a solid support.…”

Section: Aptamer Biosensor Designmentioning

confidence: 99%