Mammalian (non-human) Cell Lines
Creative Biolabs develops various stable mammalian cell lines including non-human mammalian cell lines and human cell lines for recombinant protein expression. These expression systems are able to produce post-translation modifications which closely resemble those in humans, outside of human expression systems. Various biotherapeutics including antibodies, enzymes, cytokines, hormones and clotting factors that have been licensed in the United States and Europe are produced by non-human mammalian expression systems.
Fig.1 The process to develop a mammalian cell line.1
There have been different classes of non-human stable cell lines developed to produce biotherapeutic products. The widely used non-human mammalian cell lines include CHO, mouse myeloma cell lines and rat myeloma cell lines, etc. In order to obtain high-quality stable cell lines, the most appropriate expression cassette setup, regulatory elements, targeted integration systems, selection systems, screening strategies and cloning strategies are selected and combined. Cell line and expression vector engineering facilitate the modulation of transcriptional activity to increase productivity of cell culture.
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Vectors: The key features of most vectors for stable expression include promoters, enhancers and selectable marker genes, etc. Genes of interest and selection genes can be co-expressed on the same vector with the applications of IRES elements. There are two kinds of selection systems: antibiotic selection system and metabolic selection system. The former system is based on genes which mediate resistance to antibiotic substances such as Puromycine, G418/Neomycine, and Hygromycine. While metabolic selection system uses components of metabolic key pathways as marker genes. The classical two metabolic selection marker genes are DHFR (metabolic selection system) and GS (glutamine synthetase).
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Cell lines: After the approval of tPA (tissue plasminogen activator), there are various types of mammalian cell lines that have been developed. The most widely used non-human mammalian cell lines include CHO, mouse myeloma cell lines (e.g. NS0, SP2/0), rat myeloma cell line (e.g. YB2/0), BHK, etc. Human cell line: HEK293 and its derivatives, HT-1080, PER.C6, Huh-7, etc.
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Cloning and screening technologies: Limiting dilution, colony picking and flow cytometry are methods for single cell cloning. Productivity, growth behavior and product quality attributes are the parameters that are usually analyzed to identify candidate clones.
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Creative Biolabs is capable of providing the following stable cell lines for various proteins expression:
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➢ CHO cell lines: GS system, DHFR system;
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➢ BHK cell line;
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➢ Mouse myeloma cell lines: NS0, SP2/0, etc.
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➢ Rat myeloma cell lines: YB2/0, etc.
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➢ Human cell lines: HEK293 and its derivatives, HT-1080, Huh-7, PER.C6 , etc.
Reference:
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Lai, Tingfeng, Yuansheng Yang, and Say Kong Ng. "Advances in mammalian cell line development technologies for recombinant protein production." Pharmaceuticals 6.5 (2013): 579-603. Distributed under open access license CC BY 3.0, without modification.
To discuss your Mammalian (non-human) Cell Lines demands or to request a proposal, please contact us.
For Research Use Only. Not For Clinical Use.
