Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry

Rafael Song, Michael Chan, Chris Cello, Steven R. Tannenbaum, John S. Wishnok, Jongyoon Han

    Research output: Contribution to journalArticle

    Abstract

    In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pi range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a highthroughput sample fractionation tool for targeted proteomics using LC/MS.

    Original languageEnglish (US)
    Pages (from-to)2317-2325
    Number of pages9
    JournalAnalytical Chemistry
    Volume82
    Issue number6
    DOIs
    StatePublished - Mar 15 2010

    Fingerprint

    Electrophoresis
    Sorting
    Mass spectrometry
    Peptides
    Buffers
    Proteins
    Fabrication
    Substrates
    Fractionation
    Titration
    Printing
    Salts
    Plasmas
    Glass

    ASJC Scopus subject areas

    • Analytical Chemistry

    Cite this

    Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry. / Song, Rafael; Chan, Michael; Cello, Chris; Tannenbaum, Steven R.; Wishnok, John S.; Han, Jongyoon.

    In: Analytical Chemistry, Vol. 82, No. 6, 15.03.2010, p. 2317-2325.

    Research output: Contribution to journalArticle

    Song, Rafael ; Chan, Michael ; Cello, Chris ; Tannenbaum, Steven R. ; Wishnok, John S. ; Han, Jongyoon. / Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry. In: Analytical Chemistry. 2010 ; Vol. 82, No. 6. pp. 2317-2325.
    @article{8b58042769c045fba718b87792a0fc62,
    title = "Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry",
    abstract = "In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pi range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a highthroughput sample fractionation tool for targeted proteomics using LC/MS.",
    author = "Rafael Song and Michael Chan and Chris Cello and Tannenbaum, {Steven R.} and Wishnok, {John S.} and Jongyoon Han",
    year = "2010",
    month = "3",
    day = "15",
    doi = "10.1021/ac9025219",
    language = "English (US)",
    volume = "82",
    pages = "2317--2325",
    journal = "Analytical Chemistry",
    issn = "0003-2700",
    publisher = "American Chemical Society",
    number = "6",

    }

    TY - JOUR

    T1 - Free-flow zone electrophoresis of peptides and proteins in PDMS microchip for narrow pi range sample prefractionation coupled with mass spectrometry

    AU - Song, Rafael

    AU - Chan, Michael

    AU - Cello, Chris

    AU - Tannenbaum, Steven R.

    AU - Wishnok, John S.

    AU - Han, Jongyoon

    PY - 2010/3/15

    Y1 - 2010/3/15

    N2 - In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pi range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a highthroughput sample fractionation tool for targeted proteomics using LC/MS.

    AB - In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pi range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a highthroughput sample fractionation tool for targeted proteomics using LC/MS.

    UR - http://www.scopus.com/inward/record.url?scp=77949845469&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=77949845469&partnerID=8YFLogxK

    U2 - 10.1021/ac9025219

    DO - 10.1021/ac9025219

    M3 - Article

    VL - 82

    SP - 2317

    EP - 2325

    JO - Analytical Chemistry

    JF - Analytical Chemistry

    SN - 0003-2700

    IS - 6

    ER -