Establishment of Higher Passage PER.C6 Cells for Adenovirus Manufacture

Authors

  • Marina Berdichevsky,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Search for more papers by this author
  • Marie-Pierre Gentile,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Search for more papers by this author
  • Benjamin Hughes,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Current affiliation:
    1. Biopharmaceutical Development, GlaxoSmithKline, King of Prussia, PA 19406
    Search for more papers by this author
  • Peter Meis,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Current affiliation:
    1. University of California, Berkeley, Dept. of Chemical Engineering, Berkeley, CA 94720
    Search for more papers by this author
  • Joseph Peltier,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Current affiliation:
    1. Ohio State University School of Medicine, Columbus, OH 43210
    Search for more papers by this author
  • Ilse Blumentals,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Current affiliation:
    1. Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland 4072, Australia
    Search for more papers by this author
  • John Auniņš,

    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    Search for more papers by this author
  • Nedim Emil Altaras

    Corresponding author
    1. Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486
    • Fermentation and Cell Culture, Bioprocess R&D, Merck & Co., Inc., P.O. Box 4, West Point, Pennsylvania 19486. Ph: (215) 652–0592. Fax: (215) 652–8691
    Search for more papers by this author

Abstract

PER.C6 cells, an industrially relevant cell line for adenovirus manufacture, were extensively passaged in serum-free suspension cell culture to better adapt them to process conditions. The changes in cell physiology that occurred during this passaging were characterized by investigating cell growth, cell size, metabolism, and cultivation of replication-deficient adenovirus. The changes in cell physiology occurred gradually as the population doubling level, the number of times the cell population had doubled, increased. Higher passage PER.C6 (HP PER.C6) proliferated at a specific growth rate of 0.043 h-1, 2-fold faster than lower passage PER.C6, and were capable of proliferation from lower inoculation cell densities. HP PER.C6 cell volume was 16% greater, and cellular yields on glucose, lactate, oxygen, and amino acids were greater as well. In batch cultures, HP PER.C6 cells volumetrically produced 3-fold more adenovirus, confirmed with three different constructs. The increase in productivity was also seen on a cell-specific basis. Although HP PER.C6 were more sensitive to the “cell density effect”, requiring lower infection cell densities for optimal specific productivity, they proliferated more after infection than lower passage PER.C6, increasing the number of cells available for virus production. The extensive passaging established HP PER.C6 cells with several desirable attributes for adenovirus manufacture.

Ancillary