Communication to the Editor

Integrated single-cell analysis shows Pichia pastoris secretes protein stochastically

  1. Kerry Routenberg Love1,
  2. Vasiliki Panagiotou1,
  3. Bo Jiang2,
  4. Terrance A. Stadheim2,3,
  5. J. Christopher Love1,4,*

Article first published online: 10 FEB 2010

DOI: 10.1002/bit.22688

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 106, Issue 2, pages 319–325, 1 June 2010

Additional Information

How to Cite

Love, K. R., Panagiotou, V., Jiang, B., Stadheim, T. A. and Love, J. C. (2010), Integrated single-cell analysis shows Pichia pastoris secretes protein stochastically. Biotechnol. Bioeng., 106: 319–325. doi: 10.1002/bit.22688

Author Information

  1. 1

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

  2. 2

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

  3. 3

    Strain Development Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire

  4. 4

    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.

Publication History

  1. Issue published online: 16 APR 2010
  2. Article first published online: 10 FEB 2010
  3. Accepted manuscript online: 10 FEB 2010 12:00AM EST
  4. Manuscript Accepted: 25 JAN 2010
  5. Manuscript Revised: 17 DEC 2009
  6. Manuscript Received: 16 OCT 2009

Funded by

  • Merck & Co

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Keywords:

  • yeast;
  • heterologous protein secretion;
  • Pichia pastoris;
  • clonal selection;
  • microengraving;
  • single cell

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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