Recombinant interferon-γ secreted by chinese hamster ovary-320 cells cultivated in suspension in protein-free media is protected against extracellular proteolysis by the expression of natural protease inhibitors and by the addition of plant protein hydrolysates to the culture medium
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Biosafety requirements increasingly restrict the cultivation of mammalian cells producing therapeutic glycoproteins to conditions that are devoid of any compound of animal origin. On cultivation in serum-free media, the proteases inhibitors, usually found in serum, cannot protect secreted recombinant proteins against unwanted endogenous proteolysis. Chinese hamster ovary (CHO) cells, secreting recombinant human interferon-γ (CHO-320 cell line) and cultivated in suspension in an original protein-free medium, expressed at least two members of the matrix metalloproteinases (MMP), either at the cell surface (proMMP-14 and MMP-14) or secreted (proMMP-9). In addition, tissue- and urinary-type plasminogen activators were also secreted in such culture conditions. At the cell surface, dipeptidyl peptidase IV and tripeptidyl peptidase II (TPPII) activities were also detected, and their activities decreased during time course of batch cultures. The proteolytic activities of these proteins were counterbalanced by (1) their expression as zymogens (proMMP-9, proMMP-14), (2) the expression of their natural inhibitors, tissue inhibitors of metalloproteinases-1 and-2 and plasminogen activator inhibitor-1 (PAI-1), or (3) the addition of plant protein hydrolysates to the culture medium, acting as a nonspecific source of TPPII inhibitors. This study points out that, even in protein-free media, recombinant proteins secreted by CHO cells are actively protected against physiological and unwanted extracellular proteolysis either by endogenous or by exogenous inhibitors.
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- Recombinant interferon-γ secreted by chinese hamster ovary-320 cells cultivated in suspension in protein-free media is protected against extracellular proteolysis by the expression of natural protease inhibitors and by the addition of plant protein hydrolysates to the culture medium
In Vitro Cellular & Developmental Biology - Animal
Volume 41, Issue 3-4 , pp 83-91
- Cover Date
- Print ISSN
- Online ISSN
- Additional Links
- Chinese hamster ovary cells
- matrix metalloproteinases
- plasminogen activators
- plant peptones
- Industry Sectors
- Author Affiliations
- 1. Laboratoire de Biochimie cellulaire, Institut des Sciences de la Vie (ISV) and Université catholique de Louvain, B-1348, Louvain-la-Neuve, Belgium
- 2. Laboratoire de Biologie cellulaire, Institut des Sciences de la Vie (ISV) and Université catholique de Louvain, B-1348, Louvain-la-Neuve, Belgium
- 3. Laboratoire de Chimie biologique, Université Mons-Hainaut, B-7000, Mons, Belgium
- 4. Unité de Génie biologique, Institut des Sciences de la Vie (ISV) and Université catholique de Louvain, B-1348, Louvain-la-Neuve, Belgium