Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential

Rafael Song, Stephanie Hsu, Anna L. Stevens, Jongyoon Han

    Research output: Contribution to journalArticle

    Abstract

    Efficient sample preparation tools are the key to measuring molecular signals in a complex biological system. A novel continuous-flow isoelectric point (pI)-based sorting technique has been developed for proteins and peptides in a microfluidic chip format. It can sort biomolecules at a relatively high flow rate of up to 10 μL/min and does not require carrier ampholytes, which can create molecular backgrounds for subsequent analysis. Furthermore, the electrophoretic field required to run the pI-based sorting is generated by the diffusion of buffer ions in situ, at the liquid junction between two laminar flows within the microfluidic channel. Utilizing the diffusion potential in combination with a pH difference between the buffers, we demonstrated a separation of binary mixtures of pI markers and proteins without applying any external field. The sorting resolution and its efficiency are sufficiently high for sample preparation and could be further improved by optimizing buffers or with an additional transverse electric field. Once fully developed, it can potentially be a pI-based sample fractionation tool for proteomic analysis of complex biomolecule samples.

    Original languageEnglish (US)
    Pages (from-to)3528-3536
    Number of pages9
    JournalAnalytical Chemistry
    Volume78
    Issue number11
    DOIs
    StatePublished - Jun 1 2006

    Fingerprint

    Sorting
    Microfluidics
    Buffers
    Biomolecules
    Peptides
    Ampholyte Mixtures
    Proteins
    Biological systems
    Fractionation
    Binary mixtures
    Laminar flow
    Electric fields
    Flow rate
    Ions
    Liquids

    ASJC Scopus subject areas

    • Analytical Chemistry

    Cite this

    Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential. / Song, Rafael; Hsu, Stephanie; Stevens, Anna L.; Han, Jongyoon.

    In: Analytical Chemistry, Vol. 78, No. 11, 01.06.2006, p. 3528-3536.

    Research output: Contribution to journalArticle

    Song, Rafael ; Hsu, Stephanie ; Stevens, Anna L. ; Han, Jongyoon. / Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential. In: Analytical Chemistry. 2006 ; Vol. 78, No. 11. pp. 3528-3536.
    @article{973e7ce1f5bb4ab49143824d7f6943b0,
    title = "Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential",
    abstract = "Efficient sample preparation tools are the key to measuring molecular signals in a complex biological system. A novel continuous-flow isoelectric point (pI)-based sorting technique has been developed for proteins and peptides in a microfluidic chip format. It can sort biomolecules at a relatively high flow rate of up to 10 μL/min and does not require carrier ampholytes, which can create molecular backgrounds for subsequent analysis. Furthermore, the electrophoretic field required to run the pI-based sorting is generated by the diffusion of buffer ions in situ, at the liquid junction between two laminar flows within the microfluidic channel. Utilizing the diffusion potential in combination with a pH difference between the buffers, we demonstrated a separation of binary mixtures of pI markers and proteins without applying any external field. The sorting resolution and its efficiency are sufficiently high for sample preparation and could be further improved by optimizing buffers or with an additional transverse electric field. Once fully developed, it can potentially be a pI-based sample fractionation tool for proteomic analysis of complex biomolecule samples.",
    author = "Rafael Song and Stephanie Hsu and Stevens, {Anna L.} and Jongyoon Han",
    year = "2006",
    month = "6",
    day = "1",
    doi = "10.1021/ac052156t",
    language = "English (US)",
    volume = "78",
    pages = "3528--3536",
    journal = "Analytical Chemistry",
    issn = "0003-2700",
    publisher = "American Chemical Society",
    number = "11",

    }

    TY - JOUR

    T1 - Continuous-flow pI-based sorting of proteins and peptides in a microfluidic chip using diffusion potential

    AU - Song, Rafael

    AU - Hsu, Stephanie

    AU - Stevens, Anna L.

    AU - Han, Jongyoon

    PY - 2006/6/1

    Y1 - 2006/6/1

    N2 - Efficient sample preparation tools are the key to measuring molecular signals in a complex biological system. A novel continuous-flow isoelectric point (pI)-based sorting technique has been developed for proteins and peptides in a microfluidic chip format. It can sort biomolecules at a relatively high flow rate of up to 10 μL/min and does not require carrier ampholytes, which can create molecular backgrounds for subsequent analysis. Furthermore, the electrophoretic field required to run the pI-based sorting is generated by the diffusion of buffer ions in situ, at the liquid junction between two laminar flows within the microfluidic channel. Utilizing the diffusion potential in combination with a pH difference between the buffers, we demonstrated a separation of binary mixtures of pI markers and proteins without applying any external field. The sorting resolution and its efficiency are sufficiently high for sample preparation and could be further improved by optimizing buffers or with an additional transverse electric field. Once fully developed, it can potentially be a pI-based sample fractionation tool for proteomic analysis of complex biomolecule samples.

    AB - Efficient sample preparation tools are the key to measuring molecular signals in a complex biological system. A novel continuous-flow isoelectric point (pI)-based sorting technique has been developed for proteins and peptides in a microfluidic chip format. It can sort biomolecules at a relatively high flow rate of up to 10 μL/min and does not require carrier ampholytes, which can create molecular backgrounds for subsequent analysis. Furthermore, the electrophoretic field required to run the pI-based sorting is generated by the diffusion of buffer ions in situ, at the liquid junction between two laminar flows within the microfluidic channel. Utilizing the diffusion potential in combination with a pH difference between the buffers, we demonstrated a separation of binary mixtures of pI markers and proteins without applying any external field. The sorting resolution and its efficiency are sufficiently high for sample preparation and could be further improved by optimizing buffers or with an additional transverse electric field. Once fully developed, it can potentially be a pI-based sample fractionation tool for proteomic analysis of complex biomolecule samples.

    UR - http://www.scopus.com/inward/record.url?scp=33744960152&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=33744960152&partnerID=8YFLogxK

    U2 - 10.1021/ac052156t

    DO - 10.1021/ac052156t

    M3 - Article

    VL - 78

    SP - 3528

    EP - 3536

    JO - Analytical Chemistry

    JF - Analytical Chemistry

    SN - 0003-2700

    IS - 11

    ER -