Microfluidic Library Screening for Mapping Antibody Epitopes

Bessette, Paul H.; Hu, Xiaoyuan; Soh, H. Tom; Daugherty, Patrick S.

doi:10.1021/ac0616916

Cited by 52 publications

(52 citation statements)

References 19 publications

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“…12͑a͒. The antibody fingerprints identified by Bessette et al 202 were comparable to those obtained using state-of-the-art commercial cell sorting instrumentation. This method has been extended even further 111 with the demonstration of the capability to simultaneously enrich multiple, distinct target cells into independent fractions.…”

Section: Biomedicalsupporting

confidence: 63%

“…Rare target cells were enriched by more than 200-fold in a single round of sorting at a single-channel throughput of 10 000 cells/s. Bessette et al 202 extended this methodology to develop the capability to screen molecular libraries using a disposable microfluidic device. This provides the potential to simplify and automate reagent generation and to develop integrated bioanalytical systems for clinical diagnostics.…”

Section: Biomedicalmentioning

confidence: 99%

“…This provides the potential to simplify and automate reagent generation and to develop integrated bioanalytical systems for clinical diagnostics. Antibody epitopes were mapped by Bessette et al 202 using a disposable microfluidic DEP device to screen a combinatorial peptide library composed of 5 ϫ 10 8 members displayed on bacterial cells. On-chip library screening was achieved in a two-stage, continuous-flow microfluidic sorter that separates antibody-binding target cells captured on microspheres through DEP "funneling" electrodes of the form described by Schnelle et al 69 and Fiedler et al 108 and depicted in Fig.…”

Section: Biomedicalmentioning

confidence: 99%

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Review Article—Dielectrophoresis: Status of the theory, technology, and applications

2010

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A review is presented of the present status of the theory, the developed technology and the current applications of dielectrophoresis ͑DEP͒. Over the past 10 years around 2000 publications have addressed these three aspects, and current trends suggest that the theory and technology have matured sufficiently for most effort to now be directed towards applying DEP to unmet needs in such areas as biosensors, cell therapeutics, drug discovery, medical diagnostics, microfluidics, nanoassembly, and particle filtration. The dipole approximation to describe the DEP force acting on a particle subjected to a nonuniform electric field has evolved to include multipole contributions, the perturbing effects arising from interactions with other cells and boundary surfaces, and the influence of electrical double-layer polarizations that must be considered for nanoparticles. Theoretical modelling of the electric field gradients generated by different electrode designs has also reached an advanced state. Advances in the technology include the development of sophisticated electrode designs, along with the introduction of new materials ͑e.g., silicone polymers, dry film resist͒ and methods for fabricating the electrodes and microfluidics of DEP devices ͑photo and electron beam lithography, laser ablation, thin film techniques, CMOS technology͒. Around three-quarters of the 300 or so scientific publications now being published each year on DEP are directed towards practical applications, and this is matched with an increasing number of patent applications. A summary of the US patents granted since January 2005 is given, along with an outline of the small number of perceived industrial applications ͑e.g., mineral separation, micropolishing, manipulation and dispensing of fluid droplets, manipulation and assembly of micro components͒. The technology has also advanced sufficiently for DEP to be used as a tool to manipulate nanoparticles ͑e.g., carbon nanotubes, nano wires, gold and metal oxide nanoparticles͒ for the fabrication of devices and sensors. Most efforts are now being directed towards biomedical applications, such as the spatial manipulation and selective separation/enrichment of target cells or bacteria, high-throughput molecular screening, biosensors, immunoassays, and the artificial engineering of three-dimensional cell constructs. DEP is able to manipulate and sort cells without the need for biochemical labels or other bioengineered tags, and without contact to any surfaces. This opens up potentially important applications of DEP as a tool to address an unmet need in stem cell research and therapy.

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Section: Biomedicalsupporting

confidence: 63%

Section: Biomedicalmentioning

confidence: 99%

Section: Biomedicalmentioning

confidence: 99%

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Review Article—Dielectrophoresis: Status of the theory, technology, and applications

2010

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“…In addition to the entry of nontarget cells into the collection outlets, we note that the purity performance is also reduced by cross-over of targets between the two collection outlets (i.e., entry of target 1 into outlet 2, and target 2 into outlet 1). Additional gains in purity could be achieved by implementing multiple sorting stages within a single device, as we have demonstrated with dielectrophoretic sorting devices (25). However, we believe that an important source of this cross-contamination may not originate from the device but rather from heterogeneity in the magnetic properties of the tags (26,27).…”

Section: Discussionmentioning

confidence: 89%

Multitarget magnetic activated cell sorter

Adams¹,

Kim

Soh

2008

Proc. Natl. Acad. Sci. U.S.A.

338

299

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Magnetic selection allows high-throughput sorting of target cells based on surface markers, and it is extensively used in biotechnology for a wide range of applications from in vitro diagnostics to cell-based therapies. However, existing methods can only perform separation based on a single parameter (i.e., the presence or absence of magnetization), and therefore, the simultaneous sorting of multiple targets at high levels of purity, recovery, and throughput remains a challenge. In this work, we present an alternative system, the multitarget magnetic activated cell sorter (MT-MACS), which makes use of microfluidics technology to achieve simultaneous spatially-addressable sorting of multiple target cell types in a continuous-flow manner. We used the MT-MACS device to purify 2 types of target cells, which had been labeled via target-specific affinity reagents with 2 different magnetic tags with distinct saturation magnetization and size. The device was engineered so that the combined effects of the hydrodynamic force produced from the laminar flow and the magnetophoretic force produced from patterned ferromagnetic structures within the microchannel result in the selective purification of the differentially labeled target cells into multiple independent outlets. We demonstrate here the capability to simultaneously sort multiple magnetic tags with >90% purity and >5,000-fold enrichment and multiple bacterial cell types with >90% purity and >500-fold enrichment at a throughput of 10 9 cells per hour.cell sorting ͉ magnetophoresis ͉ microfluidics ͉ separation

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“…Angled electrode structures have also been used as "funnels" to guide particle motion within a channel or as part of the function of a microsystem [55][56][57][58][59]. As DEP separators, angled electrode structures have been found favorable in their reliance on negative DEP for inducing forces of different strength on particles of different properties.…”

Section: Dielectrophoretic Separation Using Angled Electrodesmentioning

confidence: 99%

Continuous separation of colloidal particles using dielectrophoresis

2013

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Continuous separation of colloidal particles using dielectrophoresisDielectrophoresis is the movement of particles in nonuniform electric fields and has been of interest for application to manipulation and separation at and below the microscale. This technique has the advantages of being noninvasive, nondestructive, and noncontact, with the movement of particle achieved by means of electric fields generated by miniaturized electrodes and microfluidic systems. Although the majority of applications have been above the microscale, there is increasing interest in application to colloidal particles around a micron and smaller. This paper begins with a review of colloidal and nanoscale dielectrophoresis with specific attention paid to separation applications. An innovative design of integrated microelectrode array and its application to flow-through, continuous separation of colloidal particles is then presented. The details of the angled chevron microelectrode array and the test microfluidic system are then discussed. The variation in device operation with applied signal voltage is presented and discussed in terms of separation efficiency, demonstrating 99.9% separation of a mixture of colloidal latex spheres. Keywords:Colloidal / Dielectrophoresis / Separation DOI 10.1002/elps.2012004661 Introduction GeneralDielectrophoresis (DEP) is the motion arising from the interaction of nonuniform electric fields and the induced electrical dipole in polarizable particles [1][2][3][4][5]. The standard electrical technique of electrophoretic separation works when suspended charged particles migrate under the influence of applied electric field. DEP has distinct advantages over this technique in that it can separate neutral particles as well, allowing the separation of particles without having to physiologically alter them; which gives minimum particle handling and no damage. DEP also typically uses microfabricated electrodes to generate highly nonuniform electric fields, which allows it to be performed locally, whereas electrophoretic separation is generally a long-range effect acting between two large external electrodes. Due to the requirements for microfabrication, the localized nature of DEP, and the rapid growth of microfluidic Labon-a-Chip systems or TAS in the past two decades in both research and commercial sectors [6][7][8], there has been a recent growth in the application of DEP to these areas. The first practical application of Lab-on-a-Chip was in fact on-chip CE [9] and with recent advances in the synthesis and the charCorrespondence: Dr. Nicolas Green, Nano Group, School of Electronics and Computer Science, University of Southampton, Highfield, Southampton, SO17 1BJ, UK E-mail: ng2@ecs.soton.ac.uk Fax: +44-2380593029 acterization of size-selected particles in the submicron and nanometer (colloidal) range, an investigation of their physical and chemical properties is now possible [10]. The capability to produce small scale devices allows the development of entirely novel experiments and chip-based analytical system...

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Microfluidic Library Screening for Mapping Antibody Epitopes

Cited by 52 publications

References 19 publications

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

Review Article—Dielectrophoresis: Status of the theory, technology, and applications

Multitarget magnetic activated cell sorter

Continuous separation of colloidal particles using dielectrophoresis

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