Poly(oxyethylene) Based Surface Coatings for Poly(dimethylsiloxane) Microchannels

Hellmich, Wibke; Regtmeier, Jan; Duong, Ta Nguyen Binh; Ros, Robert; Anselmetti, Dario; Ros, Alexandra

doi:10.1021/la0510432

Cited by 114 publications

(121 citation statements)

References 43 publications

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“…In this study, we employed adsorptive treatment with a tri-block-copolymer (F108) to prevent non-specific protein adsorption and control EOF. 62 Recently, we could also demonstrate that dynamic coating, in which the coating agent is apparent in the buffer throughout the entire experiment is able to reduce EOF even further. 64 Hence, we employed this dynamic coating procedure with 3 mM F108 as final concentration in the buffer and studied its effect on streaming DEP of IgG.…”

Section: E Influence On Protein Dep Due To Surfactantsmentioning

confidence: 89%

“…Figures 3(a)-3(c) depict the concentration profiles for IgG (with positive DEP) in dependence of changes of l eo . We started with a typical l eo of 1.5 Â 10 À8 m 2 /Vs for oxygen plasma treated PDMS with F108 coating according to a static coating procedure 62 and varied the mobility values by up to an order of magnitude.…”

Section: B Influence Of Electroosmosismentioning

confidence: 99%

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Tuning direct current streaming dielectrophoresis of proteins

et al. 2012

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Dielectrophoresis (DEP) of biomolecules has large potential to serve as a novel selectivity parameter for bioanalytical methods such as (pre)concentration, fractionation, and separation. However, in contrast to well-characterized biological cells and (nano)particles, the mechanism of protein DEP is poorly understood, limiting bioanalytical applications for proteins. Here, we demonstrate a detailed investigation of factors influencing DEP of diagnostically relevant immunoglobulin G (IgG) molecules using insulator-based DEP (iDEP) under DC conditions. We found that the pH range in which concentration of IgG due to streaming iDEP occurs without aggregate formation matches the pH range suitable for immunoreactions. Numerical simulations of the electrokinetic factors pertaining to DEP streaming in this range further suggested that the protein charge and electroosmotic flow significantly influence iDEP streaming. These predictions are in accordance with the experimentally observed pH-dependent iDEP streaming profiles as well as the determined IgG molecular properties. Moreover, we observed a transition in the streaming behavior caused by a change from positive to negative DEP induced through micelle formation for the first time experimentally, which is in excellent qualitative agreement with numerical simulations. Our study thus relates molecular immunoglobulin properties to observed iDEP, which will be useful for the future development of protein (pre)concentration or separation methods based on DEP.

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Section: E Influence On Protein Dep Due To Surfactantsmentioning

confidence: 89%

Section: B Influence Of Electroosmosismentioning

confidence: 99%

Tuning direct current streaming dielectrophoresis of proteins

et al. 2012

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“…Liquid connections were incorporated by punching holes with biopsy punchers (WPI). To preventexcessive sticking of silk proteins to the walls of the microfluidic device, a 1% wt/vol solution of F108 block copolymer was introduced into the channels and incubated for 1 h at 60°C (26). Before use, all channels were thoroughly rinsed with 10 mM Tris buffer, pH 8.…”

Section: Methodsmentioning

confidence: 99%

Assembly mechanism of recombinant spider silk proteins

Rammensee¹,

Slotta

Scheibel

et al. 2008

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

356

377

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Spider silk threads are formed by the irreversible aggregation of silk proteins in a spinning duct with dimensions of only a few micrometers. Here, we present a microfluidic device in which engineered and recombinantly produced spider dragline silk proteins eADF3 (engineered Araneus diadematus fibroin) and eADF4 are assembled into fibers. Our approach allows the direct observation and identification of the essential parameters of dragline silk assembly. Changes in ionic conditions and pH result in aggregation of the two proteins. Assembly of eADF3 fibers was induced only in the presence of an elongational flow component. Strikingly, eADF4 formed fibers only in combination with eADF3. On the basis of these results, we propose a model for dragline silk aggregation and early steps of fiber assembly in the microscopic regime.colloids ͉ microfluidics ͉ protein materials ͉ rheology

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“…A significant improvement of the detection limit should arise by controlling PDMS protein interaction to circumvent unspecific adhesion. For that reason we are currently investigating PDMS surface coatings with poly(oxyethylene) compounds [37].…”

Section: Protein Separation and Uv-lif Detectionmentioning

confidence: 99%

Single cell manipulation, analytics, and label-free protein detection in microfluidic devices for systems nanobiology

et al. 2005

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Single cell analytics for proteomic analysis is considered a key method in the framework of systems nanobiology which allows a novel proteomics without being subjected to ensemble-averaging, cell-cycle, or cell-population effects. We are currently developing a single cell analytical method for protein fingerprinting combining a structured microfluidic device with latest optical laser technology for single cell manipulation (trapping and steering), free-solution electrophoretical protein separation, and (label-free) protein detection. In this paper we report on first results of this novel analytical device focusing on three main issues. First, single biological cells were trapped, injected, steered, and deposited by means of optical tweezers in a poly(dimethylsiloxane) microfluidic device and consecutively lysed with SDS at a predefined position. Second, separation and detection of fluorescent dyes, amino acids, and proteins were achieved with LIF detection in the visible (VIS) (488 nm) as well as in the deep UV (266 nm) spectral range for label-free, native protein detection. Minute concentrations of 100 fM injected fluorescein could be detected in the VIS and a first protein separation and label-free detection could be achieved in the UV spectral range. Third, first analytical experiments with single Sf9 insect cells (Spodoptera frugiperda) in a tailored microfluidic device exhibiting distinct electropherograms of a green fluorescent protein-construct proved the validity of the concept. Thus, the presented microfluidic concept allows novel and fascinating single cell experiments for systems nanobiology in the future.

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Poly(oxyethylene) Based Surface Coatings for Poly(dimethylsiloxane) Microchannels

Cited by 114 publications

References 43 publications

Tuning direct current streaming dielectrophoresis of proteins

Tuning direct current streaming dielectrophoresis of proteins

Assembly mechanism of recombinant spider silk proteins

Single cell manipulation, analytics, and label-free protein detection in microfluidic devices for systems nanobiology

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