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Integrated single‐cell analysis shows Pichia pastoris secretes protein stochastically |
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| Authors: | Kerry Routenberg Love Vasiliki Panagiotou Bo Jiang Terrance A. Stadheim J. Christopher Love |
| Affiliation: | 1. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, Massachusetts 02139;2. telephone: 617‐324‐2300;3. fax: 617‐258‐5042;4. High Throughput Screening & Strain Selection Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire;5. Strain Development Group, GlycoFi, Inc. (A Wholly Owned Subsidiary of Merck & Co.) Lebanon, New Hampshire;6. The Eli and Edythe L. Broad Institute, 7 Cambridge Center, Cambridge, Massachusetts 02139 |
| 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. |
| Keywords: | yeast heterologous protein secretion Pichia pastoris clonal selection microengraving single cell |