サイトメトリーを利用した迅速なCHO細胞タンパク質発現・精製システム A rapid protein expression and purification system in Chinese hamster ovary cells using flow cytometry

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著者

    • 鈴木 淳 SUZUKI Jun
    • 奈良先端科学技術大学院大学 バイオサイエンス研究科細胞増殖学講座 Laboratory of Molecular Oncology, Nara Institute of Science and Technology
    • 圓岡 真宏 MARUOKA Masahiro
    • 奈良先端科学技術大学院大学 バイオサイエンス研究科細胞増殖学講座 Laboratory of Molecular Oncology, Nara Institute of Science and Technology
    • 川田 滋久 KAWATA Shigehisa
    • 奈良先端科学技術大学院大学 バイオサイエンス研究科細胞増殖学講座 Laboratory of Molecular Oncology, Nara Institute of Science and Technology
    • 宍戸 知行 SHISHIDO Tomoyuki
    • 奈良先端科学技術大学院大学 バイオサイエンス研究科細胞増殖学講座 Laboratory of Molecular Oncology, Nara Institute of Science and Technology

抄録

<p>Chinese hamster ovary (CHO) cell is widely adopted in industry for production of desired proteins such as monoclonal antibodies, hormones and interferon. However, stable expression of a desired protein in CHO cells requires a time-consuming step of antibiotic resistance-based cell cloning. Recently, we have developed a retrovirus-based system for stable expression of a desired protein in the CHO-S cell, a derivative of CHO cell. Although CHO cells are resistant to retrovirus infection, stable expression of ecotropic retrovirus receptor (EcoR) in those cells enabled infection. We constructed the retroviral vector pGX GFP, which expresses the desired protein as a GST fusion construct and the GFP as a marker protein. Multiple infection of retrovirus helped to increase the amount of the desired protein in single cells, and collection of GFP-high cell population by flow cytometry facilitated to obtain cell population expressing the desired protein at high levels. This system made it possible to obtain the reasonable amount of the desired protein (100µg) in about two weeks. In this review, we will discuss the advantage and the point of issue of this system.</p>

収録刊行物

  • Cytometry research = 日本サイトメトリー学会機関誌

    Cytometry research = 日本サイトメトリー学会機関誌 17(1), 35-42, 2007-03-25

    日本サイトメトリー学会

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