CHO-K1安定細胞株作製 CHO-K1安定細胞株作製

CHO-K1安定細胞株作製

CHO-K1 Stable Cell line Generation
  • 高収量の保証
  • 10年以上の経験を有した研究者のサポート
  • 商業用サブライセンスの可能

サービス概要

細胞株の生成は、医薬品開発プロセスにおいて重要な役割を果たします。産業の要求を満たすリコンビナントタンパク質や抗体のための細胞株を効率的に生産することは、貴重な時間を節約し、総合的なコストを削減することができます。


Biointronでは、ECACCライセンスの付与を受けたCHO-K1細胞を用い、専門的な細胞株開発サービスを提供しています。経験豊富なチームが、高収量の細胞株を迅速に作成し、お客様の後工程のCMC開発ニーズに合わせてカスタマイズします。

CHO-K1安定細胞株作製サービス概要

ハイライト

高収量の保証

  • 抗体生産において5グラム/リットル以上の高収量を保証します。
  • 細胞とプロセスを最適化しました。
  • 迅速な対応で、最短3ヶ月の納期を実現します。

10年以上の経験を有した研究者のサポート

  • 10年以上にわたる安定した細胞株開発の実績があります。
  • 100以上の高収量細胞株の成功した提供実績がございます。
  • お客様の特定のニーズに合わせた効率的で合理化されたプロセスをご提供します。

商業用サブライセンスの可能

  • 研究用品質または商業用品質の株を提供しております。
  • 商業サブライセンスも提供されています。

CHO-K1安定細胞株作製
サービスの詳細

ステップ 詳細 作業時間 納品
細胞株開発
遺伝子合成と一過性発現
  • コドン最適化と遺伝子合成
  • 発現ベクターへのサブクローニング
  • 一時的な発現
2-3週間
  • コドン最適化と遺伝子合成
  • 発現ベクターへのサブクローニング
安定細胞プールを作成
  • 安定トランスフェクション
  • 安定細胞プールの選択と増幅
8-9週間
シグナルクローンの選択とスクリーニング
  • クローンのサブクローニング
  • クローンの特性評価
8-9週間
  • 安定プールから回収された50ml上清
  • レポート
プライマリー細胞バンクの作成
  • プライマリーセルバンク(PCB)の調製
  • PCB安定性試験
~3ヵ月
  • プライマリー細胞バンク-マイコプラズマ検査済み
  • 分析証明書
  • レポート
CHO-K1細胞レポート
  • CHO-K1細胞株の詳細レポートとドキュメント
N/A
  • CHO-K1細胞株のレポートおよび資料
CHO-K1細胞サブライセンスサービス
CHO-K1細胞サブライセンス
  • CHO-K1細胞株の詳細レポートとドキュメント
  • CHO-K1サブライセンス契約
3-5 days
  • 2チューブ CHO-K1 懸濁細胞
  • CHO-K1細胞株のレポートおよび資料
  • CHO-K1サブライセンス契約書

事例研究

  • Case 1: Tag free vaccine protein
    Single cell image system is applied to confirm the monoclonality. The yield of the final obtained clone A is 2.31 g/L.
    Day 0
    Day 0
    Day 1
    Day 1
    Day 2
    Day 2
    Day 4
    Day 4
    Day 7
    Day 7
    Day 10
    Day 10
    Finally
    The stability of clone A is also tested by assessing the doubling time of the cells, cell density and yield.
    Stability Studies
    Subcultured in CD04 medium containing 25 uM MSX the doubling time:22 ± 1hours (Average: 22.6h)
    Feed batch
    Cells from passages 3, 8, 13, 18, and 23 were collected, and the cell density was assessed. The results indicated a consistent level of cell growth.
    Doubling Time of N108-54
    Cells from passages 3, 8, 13, 18, and 23 were collected, and the yield was assessed. The results indicated that over 90% productivity titer was retained for over 22 passages.
  • Case 2: Anti-PD-1
    Three single clones were selected, and their yield was assessed using three different commercial media. The results are as follows:
    Subclone Medium A (g/L) Medium B (g/L) Medium C (g/L)
    414203-N42-6-N1 3.19 5.5 2.63
    414203-N42-7-N1 6.48 2.78 1.9
    414209-N10-6-N1 7.82 9.02 7.72
    Both reducing and non-reducing CE-SDS analyses were performed to assess the purity of these three clones after one step of affinity purification. The results indicated >97% purity.
    R-CE-SDS
    Sample Name main peak % LMW % Total %
    Positive control 99.1 0.9 100.0
    B414203-N42-6-N1 98.8 1.3 100.0
    B414203-N42-7-N1 98.5 1.5 100.0
    B414209-N10-6-N1 97.8 2.2 100.0
    NR-CE-SDS
    Sample Name main peak % LMW % HMW % Total %
    Positive control 98.3 1.7 0.0 100.0
    B414203-N42-6-N1 94.6 5.4 0.0 100.0
    B414203-N42-7-N1 94.1 5.9 0.0 100.0
    B414209-N10-6-N1 93.3 6.4 0.3 100.0
    CEX-HPLC analyses were performed to assess the charge of these three clones. The results indicated a similar main component% compared to the positive control for the first two clones. Additionally, the first clone (B414203-N42-6-N1) shows the most similar basic component%.
    CEX-HPLC
    Sample Name Acidic component % Main component % Basic component % Total %
    Positive control 19.5 64.1 16.4 100.0
    B414203-N42-6-N1 16.0 69.7 14.3 100.0
    B414203-N42-7-N1 17.4 69.2 13.4 100.0
    B414209-N10-6-N1 18.1 44.6 37.3 100.0
    Currently, we have sublicensed the CHOK1BN cell line to dozens of customers, and the status of some customers' projects is as follows:
    Customer Type CLD PD Pilot Nonclinical IND Phase I Phase II
    1 Customer A ADC
    2 Customer G R-vaccines
    3 Customer H Mab
    4 Customer B Mab
    5 Customer C Fab
    6 Customer J R-vaccines
    7 Customer F R-glycoprotein
“With ECACC-licensed CHO-K1 cells, we can generate research grade or commercial grade stable cell lines, with the option for a one-time sublicensing fee for a perpetual and irrevocable license.”
Jing Cheng
Jing Cheng
Cell Line Generation Team

FAQs

  • What are CHO-K1 cell lines?

    Chinese hamster ovary (CHO) cell lines are derived from the ovary of adult, female Chinese hamsters. CHO cells were first established in 1957 by T. Puck, and were subsequently multiplied and optimized in vitro, allowing it to be cultured indefinitely. The CHO-K1 cell line was derived as a subclone from that parental CHO cell line. They are typically the preferred host expression system for recombinant antibodies due to their advantages in producing complex therapeutics and manufacturing adaptability.1

  • Why use CHO cells?

    One of the key advantages of CHO cells is their ability to produce complex proteins that are human-compatible, with post-translational modifications, such as glycosylation. This feature increases therapeutic efficacy, protein longevity, and reduces safety concerns. Furthermore, CHO cells are highly stable, having a high tolerance for fluctuations in temperature, acidity (pH), oxygen, and cell density. This adaptability makes them suitable for large-scale manufacturing, and the product can be delivered in up to several grams per liter.2

  • Stable vs. Transient Expression

    Stable expression involves the integration of the gene of interest into the genome of the host cell line, allowing for long-term, heritable expression. This is typically achieved by using viral vectors or plasmids containing the gene of interest and a selectable marker (e.g., antibiotic resistance gene) to facilitate the selection and isolation of cells that have successfully integrated the gene.

    Transient expression involves the direct introduction of the gene of interest into the host cells without integration into the genome. This is usually achieved through methods like transfection or electroporation.

    The use of either depends on your research goals. Typically, for long-term studies or consistent gene expression across multiple experiments, stable expression is preferred. But for quick screening or short-term experiments, transient expression can be more suitable.

  • Why are CHO cells used for therapeutic antibody production?

    There is an increasing number of recombinant antibodies being developed as therapeutic agents, with about 40 new antibody molecules undergoing clinical trials every year, most of which are produced from CHO cells.3,4 For instance, TZ mAb is a therapeutic antibody that was expressed by CHO cells and is specific for the human epidermal growth factor receptor 2 (HER2).

参考文献一覧

  • Fischer, S., Handrick, R., & Otte, K. (2015). The art of CHO cell engineering: A comprehensive retrospect and future perspectives. Biotechnology Advances, 33(8), 1878-1896. https://doi.org/10.1016/j.biotechadv.2015.10.015
  • Xu, X., Nagarajan, H., Lewis, N. E., Pan, S., Cai, Z., Liu, X., Chen, W., Xie, M., Wang, W., Hammond, S., Andersen, M. R., Neff, N., Passarelli, B., Koh, W., Fan, H. C., Wang, J., Gui, Y., Lee, K. H., Betenbaugh, M. J., . . . Wang, J. (2011). The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line. Nature Biotechnology, 29(8), 735-741. https://doi.org/10.1038/nbt.1932
  • Li, F., Vijayasankaran, N., Kiss, R., & Amanullah, A. (2010). Cell culture processes for monoclonal antibody production. MAbs, 2(5), 466-477. https://doi.org/10.4161/mabs.2.5.12720
  • Zhang, J., Shan, L., Liang, F., Du, C., & Li, J. (2022). Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells. Frontiers in Bioengineering and Biotechnology, 10, 856049. https://doi.org/10.3389/fbioe.2022.856049

カスタムサービス&プロダクト品

お客様の利便性を向上させるためにクッキーを使用しています。詳しくは プライバシーポリシー をご覧ください。