Get access

Proteomic Analysis for the Assessment of Different Lots of Fetal Bovine Serum as a Raw Material for Cell Culture. Part IV. Application of Proteomics to the Manufacture of Biological Drugs

Authors

  • Xiaoyang Zheng,

    1. Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, 341 Mugar Hall, 360 Huntington Avenue, Boston, Massachusetts 02115
    Search for more papers by this author
  • Haven Baker,

    1. Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, 341 Mugar Hall, 360 Huntington Avenue, Boston, Massachusetts 02115
    Search for more papers by this author
  • William S. Hancock,

    Corresponding author
    1. Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, 341 Mugar Hall, 360 Huntington Avenue, Boston, Massachusetts 02115
    • Berlex Biosciences, 2600 Hilltop Dr., Richmond, California 94804. Ph: 617–373-4818. Fax: 617–373-2855
    Search for more papers by this author
  • Farah Fawaz,

    1. Berlex Biosciences, 2600 Hilltop Dr., Richmond, California 94804
    Search for more papers by this author
  • Michael McCaman,

    1. Berlex Biosciences, 2600 Hilltop Dr., Richmond, California 94804
    Search for more papers by this author
  • Erno Pungor Jr.

    Corresponding author
    1. Berlex Biosciences, 2600 Hilltop Dr., Richmond, California 94804
    • Berlex Biosciences, 2600 Hilltop Dr., Richmond, California 94804. Ph: 617–373-4818. Fax: 617–373-2855
    Search for more papers by this author

  • For the first three papers in this series see refs 1–3

Abstract

Fetal bovine serum (FBS) is the most widely used growth supplement for cell cultures, primarily because of its high levels of growth stimulatory factors and low levels of growth inhibitory factors. Maintaining successful and consistent cell fermentations can be difficult, as FBS is a complex natural product and may vary from lot to lot even from a single manufacturer. The quality and concentration of both bulk and specific proteins can affect cell growth. Quality control tools for FBS are relatively primitive and expensive given the complexity of the sample and the large amounts of FBS used. We undertook this study to examine whether proteomics could be used as a tool to analyze the variability of different fermentation processes. We hypothesized that inconsistent cell growth in fermentations could be due to the quality of FBS and that different lots of FBS had varying concentrations of proteins such as growth stimulatory factors, growth inhibitory factors, and/or other proteins that may correlate with cellular growth rate. To investigate whether this was the case, we grew three batches of adult retinal pigment epithelial cells (ARPE-19) using three different lots of fetal bovine serum (FBS-Ia, FBS-Ib, and FBS-II). We found that the growth rate of the culture was significantly and consistently higher in the FBS-II lot. To determine why the other lots promoted different growth properties, we used proteomic techniques to analyze the protein composition of the three lots. We then performed a time course study to monitor specific changes in individual proteins in the fermentation medium. The amount of several extracellular matrix and structural proteins, which are indicators of cell growth, increased over time. Alternatively, components supplied by the FBS addition, such as nutritional-related and cell-spreading-related proteins, decreased over time.

Get access to the full text of this article

Ancillary