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Integrated single-cell analysis shows Pichia pastoris secretes protein stochastically

Authors

  • Kerry Routenberg Love,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139; telephone: 617-324-2300; fax: 617-258-5042
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  • Vasiliki Panagiotou,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139; telephone: 617-324-2300; fax: 617-258-5042
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  • Bo Jiang,

    1. High Throughput Screening & Strain Selection Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire
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  • Terrance A. Stadheim,

    1. High Throughput Screening & Strain Selection Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire
    2. Strain Development Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire
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  • J. Christopher Love

    Corresponding author
    1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139; telephone: 617-324-2300; fax: 617-258-5042
    2. The Eli and Edythe L. Broad Institute, 7 Cambridge Center, Cambridge, Massachusetts 02139
    • Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139; telephone: 617-324-2300; fax: 617-258-5042.
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Abstract

The production of heterologous proteins by secretion from cellular hosts is an important determinant for the cost of biotherapeutics. A single-cell analytical method called microengraving was used to examine the heterogeneity in secretion by the methylotrophic yeast Pichia pastoris. We show that constitutive secretion of a human Fc fragment by P. pastoris is not cell-cycle dependent, but rather fluctuates between states of high and low productivity in a stochastic manner. Biotechnol. Bioeng. 2010;106: 319–325. © 2010 Wiley Periodicals, Inc.

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