Microfluidic Free-Flow Electrophoresis for Proteomics-on-a-Chip

Dietrich Kohlheyer

doi:10.3990/1.9789036526661

Microfluidic Free-Flow Electrophoresis for Proteomics-on-a-Chip

Dietrich Kohlheyer

Research output: Thesis › PhD Thesis - Research UT, graduation UT

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Abstract

A new free-flow electrophoresis microchip with integrated permeable membranes was developed, and different substances were separated by free-flow zone electrophoresis, free-flow isoelectric focusing and free-flow field step electrophoresis. This chip contained a new type of membranes enabling a stable carrier flow with a perpendicular electrical current. Due to this chip configuration, the device performance and efficiency were superior to recently published alternative systems in terms of separation resolution and sample capacity. The results furthermore indicate that even better results are possible. Analytes were separated and focused within hundreds of milliseconds whereby only nanoliters of samples were consumed. In addition, a new sample steering method was demonstrated during free-flow zone electrophoresis, allowing the specific sorting of various components. As an alternative, a free-flow electrophoresis chip was developed with integrated platinum electrodes, whereby the generation of gas bubbles caused by electrolysis was successfully suppressed by chemical means. Gas bubbles generated by electrolysis are major concern in free-flow electrophoresis systems in general leading to distorted separation. Based on the results, a fourth free-flow chip was developed with an integrated surface plasmon resonance gold detection region. Although fabrication was successful, certain hurdles, in particular surface chemistry issues still remain to be overcome to perform separation and real-time detection of biological samples within this hyphenated micro device. A strategy for proteomics-on-a-chip was developed aiming at the separation of antigens that play a role in autoimmune diseases. In addition two new continuous flow microfluidic chips were developed allowing for continuous biochemical reactions of surface patterning applications. These devices could be of further interest in future, in particular in more complex analytical systems related to proteomics-on-a-chip.

Original language	English
Qualification	Doctor of Philosophy
Awarding Institution	University of Twente
Supervisors/Advisors	van den Berg, Albert, Supervisor Schasfoort, R.B.M. , Co-Supervisor
Thesis sponsors	Dutch Technology Foundation (STW)
Award date	20 Jun 2008
Place of Publication	Enschede
Publisher	University of Twente
Print ISBNs	978-90-365-2666-1
DOIs	https://doi.org/10.3990/1.9789036526661
Publication status	Published - 20 Jun 2008

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Thesis D. KohlheyerFinal published version, 22 MB

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title = "Microfluidic Free-Flow Electrophoresis for Proteomics-on-a-Chip",

abstract = "A new free-flow electrophoresis microchip with integrated permeable membranes was developed, and different substances were separated by free-flow zone electrophoresis, free-flow isoelectric focusing and free-flow field step electrophoresis. This chip contained a new type of membranes enabling a stable carrier flow with a perpendicular electrical current. Due to this chip configuration, the device performance and efficiency were superior to recently published alternative systems in terms of separation resolution and sample capacity. The results furthermore indicate that even better results are possible. Analytes were separated and focused within hundreds of milliseconds whereby only nanoliters of samples were consumed. In addition, a new sample steering method was demonstrated during free-flow zone electrophoresis, allowing the specific sorting of various components. As an alternative, a free-flow electrophoresis chip was developed with integrated platinum electrodes, whereby the generation of gas bubbles caused by electrolysis was successfully suppressed by chemical means. Gas bubbles generated by electrolysis are major concern in free-flow electrophoresis systems in general leading to distorted separation. Based on the results, a fourth free-flow chip was developed with an integrated surface plasmon resonance gold detection region. Although fabrication was successful, certain hurdles, in particular surface chemistry issues still remain to be overcome to perform separation and real-time detection of biological samples within this hyphenated micro device. A strategy for proteomics-on-a-chip was developed aiming at the separation of antigens that play a role in autoimmune diseases. In addition two new continuous flow microfluidic chips were developed allowing for continuous biochemical reactions of surface patterning applications. These devices could be of further interest in future, in particular in more complex analytical systems related to proteomics-on-a-chip.",

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AB - A new free-flow electrophoresis microchip with integrated permeable membranes was developed, and different substances were separated by free-flow zone electrophoresis, free-flow isoelectric focusing and free-flow field step electrophoresis. This chip contained a new type of membranes enabling a stable carrier flow with a perpendicular electrical current. Due to this chip configuration, the device performance and efficiency were superior to recently published alternative systems in terms of separation resolution and sample capacity. The results furthermore indicate that even better results are possible. Analytes were separated and focused within hundreds of milliseconds whereby only nanoliters of samples were consumed. In addition, a new sample steering method was demonstrated during free-flow zone electrophoresis, allowing the specific sorting of various components. As an alternative, a free-flow electrophoresis chip was developed with integrated platinum electrodes, whereby the generation of gas bubbles caused by electrolysis was successfully suppressed by chemical means. Gas bubbles generated by electrolysis are major concern in free-flow electrophoresis systems in general leading to distorted separation. Based on the results, a fourth free-flow chip was developed with an integrated surface plasmon resonance gold detection region. Although fabrication was successful, certain hurdles, in particular surface chemistry issues still remain to be overcome to perform separation and real-time detection of biological samples within this hyphenated micro device. A strategy for proteomics-on-a-chip was developed aiming at the separation of antigens that play a role in autoimmune diseases. In addition two new continuous flow microfluidic chips were developed allowing for continuous biochemical reactions of surface patterning applications. These devices could be of further interest in future, in particular in more complex analytical systems related to proteomics-on-a-chip.

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DO - 10.3990/1.9789036526661

M3 - PhD Thesis - Research UT, graduation UT

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PB - University of Twente

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ER -

Microfluidic Free-Flow Electrophoresis for Proteomics-on-a-Chip

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