Superporous agarose beads as a solid support for microfluidic immunoassay

Yang, Yoonsun; Nam, Soonkeon; Lee, Nae Yoon; Kim, Youn Sang; Park, Sungsu

doi:10.1016/j.ultramic.2008.04.044

Cited by 19 publications

(15 citation statements)

References 15 publications

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“…Delamarche et al first introduced the use of microfluidic channel networks for surface patterning of immunoglobulin in high throughput, parallel immunoassays (44). It should also be noted that microbeads have often been used in microfluidic systems for heterogeneous immunoassays (36,(45)(46)(47)(48) (49). In this study, the porosity of the beads is important since it lowers the fluidic resistance and increases the active surface area, enhancing assay sensitivity.…”

Section: Protein Functionalitymentioning

confidence: 92%

Recent advances in microfluidic technologies for biochemistry and molecular biology

Cho¹,

Kang²,

Choo³

et al. 2011

BMB Reports

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Section: Protein Functionalitymentioning

confidence: 92%

Recent advances in microfluidic technologies for biochemistry and molecular biology

Cho¹,

Kang²,

Choo³

et al. 2011

BMB Reports

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“…[18][19][20]23,24,46,94 The swelling property of hydrogels allows integration of actuators such as valves, allowing integration of sophisticated fluid handling functions. 133 A wide range of immobilization methods are available to hydrogels, including copolymerization of proteins, 18,19,23,94,132 activation for covalent linking of proteins, 134 or electrostatic capture on charged hydrogel. 20,24 Even with a 3-D microchannel-filling hydrogel, the microchannel surface should be functionalized for covalent anchoring of the hydrogel structure within the channel, so that the gel will not drift out of the channel under hydrodynamic pressure or applied electric field.…”

Section: Hydrogelmentioning

confidence: 99%

“…Instead of using GA reagent, Yang et al used in situ generation of aldehyde groups in macroporous agarose beads in order to covalently attach protein A. 134 Macropores (average pore size of 28 lm) in the bead can improve hydrodynamic mass transport through packed agarose beads, so a high flow rate can be used in chromatography without a large backpressure. Agarose beads were hydrolyzed in 0.2 M HCl at 55 C to form aldehyde groups.…”

Section: Covalent Immobilization-bioaffinity Immobilizationmentioning

confidence: 99%

Protein immobilization techniques for microfluidic assays

2013

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Microfluidic systems have shown unequivocal performance improvements over conventional bench-top assays across a range of performance metrics. For example, specific advances have been made in reagent consumption, throughput, integration of multiple assay steps, assay automation, and multiplexing capability. For heterogeneous systems, controlled immobilization of reactants is essential for reliable, sensitive detection of analytes. In most cases, protein immobilization densities are maximized, while native activity and conformation are maintained. Immobilization methods and chemistries vary significantly depending on immobilization surface, protein properties, and specific assay goals. In this review, we present trade-offs considerations for common immobilization surface materials. We overview immobilization methods and chemistries, and discuss studies exemplar of key approaches—here with a specific emphasis on immunoassays and enzymatic reactors. Recent “smart immobilization” methods including the use of light, electrochemical, thermal, and chemical stimuli to attach and detach proteins on demand with precise spatial control are highlighted. Spatially encoded protein immobilization using DNA hybridization for multiplexed assays and reversible protein immobilization surfaces for repeatable assay are introduced as immobilization methods. We also describe multifunctional surface coatings that can perform tasks that were, until recently, relegated to multiple functional coatings. We consider the microfluidics literature from 1997 to present and close with a perspective on future approaches to protein immobilization.

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“…While it took a significant amount of time in our experiments for the QDot target to access receptor sites deep within the bead, introducing bead pulsation (Thompson and Bau 2011) or employing smaller target molecules with higher diffusion coefficients, homogeneous agarose beads with larger pores (e.g., 2 or 4% beads), or superporous agarose beads (Gustavsson and Larsson 1996;Yang et al 2008) could significantly enhance the rate of binding. Furthermore, one may not need to wait until the target reaches receptor sites deep within the bead to achieve a detectable signal.…”

Section: Theoretical Predictionsmentioning

confidence: 99%

Porous bead-based microfluidic assay: theory and confocal microscope imaging

Thompson

Bau

2011

Microfluid Nanofluid

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Functionalized, porous microbeads provide large surface area to volume ratios, allowing the capture of relatively large quantities of target molecules from complex solutions and, thus, facilitating high-sensitivity assays. While in recent years the interest in bead-based assays has been growing, only a few studies focus on mass transfer in the bead's interior and on the binding kinetics of functionalized, porous beads. In this study, streptavidin-coated, porous agarose beads are controllably positioned within a microfluidic conduit. Biotinylated quantum dots are pumped through the conduit and used as labels to monitor target analyte binding to the beads. Confocal microscopy techniques are employed to image the concentration of the bound quantum labels as a function of position and time. Threedimensional, finite element simulations are carried out to model the mass transfer and binding kinetics within the beads. Key thermophysical properties, such as the reduced diffusivity of the quantum dots in the agarose matrix, are determined experimentally. Experimental observations are critically compared and favorably agree with theoretical predictions. The theoretical models provide a useful tool to better our understanding of the phenomena involved and to predict how various parameters affect microbead reaction kinetics and biosensor performance.

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Superporous agarose beads as a solid support for microfluidic immunoassay

Cited by 19 publications

References 15 publications

Recent advances in microfluidic technologies for biochemistry and molecular biology

Recent advances in microfluidic technologies for biochemistry and molecular biology

Protein immobilization techniques for microfluidic assays

Porous bead-based microfluidic assay: theory and confocal microscope imaging

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