Construction of an Arty apoptosis CHOCell Line for Biopharmaceutical Production
2003; Elsevier BV; Volume: 19; Issue: 3 Linguagem: Inglês
ISSN
1872-2075
AutoresLai Da Zhi, Ling Fu, Yu Ming, Qi Lian Quan, Weng Shao Jie, Ting Yu, Haitao Wang, Wei Chen,
Tópico(s)Virus-based gene therapy research
ResumoMammalian cells are prone to apoptosis when cultured in large scale for production of biopharmaceuticals .And this will reduce production duration and result in high cost of production .Apoptosis is triggered by various factors，and delicately re- gulated by a set of genes . Bcl- 2，a component integrated in mitochondria membrane，is an important member of these genes.by maintaining the integrity of mitochondria membrane，Bcl-2 keeps cytochrome C from releasing into cytoplasm，and thus blocks the activation of caspases，and subsequent onset of apoptosis.Over-expression of Bcl-2 has proven to be useful in blocking apoptosis in various cell lines，including CHO，hybridoma，myeloma，lymphoma and insect cells .Ammonia，a metabolite of culturedcells，however，showed apparent pro-apoptosis activity.In living cells，ammonia can be utilized by glutamine synthetase(GS) to synthesize glutamine，and thus lower the concentration of ammonia in medium，and its negative effects.Glutamine is essential to living cells .If not added into medium，glutamine can only be synthesized by GS，which makes GS a qualified selectionmarker .This marker can be used for gene amplification by adding into medium increased concentration of MSX, an inhibitor of GS .In this study ，、over-expressed Bcl-2 using GS amplification in a recombinant CHO cell line stably expressing human inter fe roir [3.The modified cell line，with higher expression of Bcl-2 and lower production of ammonia，exhibited good anti apoptosis quality and higher interferoiro production in continuous culture.
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