亚洲国产日本_互换娇妻爽文100系列电影_26uuu精品一区二区_亚洲一区 中文字幕 久久_欧美亚洲日本在线观看成_潮喷中出日韩中文字幕_流白浆视频网站_91丨永久精品丨人妖_国产成?人片在线观看视频_国产又粗又猛又爽免费视频麻豆

CHO-K1穩定細胞株構建 CHO-K1穩定細胞株構建
細胞株

CHO-K1穩定細胞株構建

  • 高表達量
  • 經驗豐富
  • CHO-K1細胞株亞授權

服務介紹

細胞株構建在抗體藥物開發過程中起著至關重要的作用。細胞株的高效產能，可以為工業生產重組蛋白和抗體節省寶貴的時間并降低總體成本。

百英可以提供直接用于CMC大規模生產的穩定細胞株開發服務，我們的CHO K1細胞株來源清晰，表達量高，來源ECACC，可以全球亞授權。

CHO-K1穩定細胞株構建 Overview

服務亮點

高表達量

  • 抗體表達量>5g/L
  • 工藝完善
  • 細胞池周期2-2.5個月，單克隆4個月

經驗豐富

  • 10年以上細胞株開發經驗
  • 上百個細胞株項目開發
  • 根據需求，量身定做構建方案

CHO K1細胞株亞授權

  • 商業化亞授權
  • 根據項目授權
  • 一次性授權
立即咨詢

CHO-K1穩定細胞株構建
服務詳情

服務步驟 服務詳情 周期 交付物
基因合成、亞克隆及表達純化
  • 密碼子優化和基因合成
  • 亞克隆到表達質粒
  • 質粒驗證、大量抽提及表達純化
 2-3周
  • 分子克隆及表達載體報告
  • 表達載體構建報告
  • 表達載體測序報告
細胞池構建
  • 穩定轉染
  • 細胞池篩選
 6-7周
  • 轉染載體Minipool報告
  • 亞克隆篩選報告
單克隆篩選
  • 亞克隆
  • 單克隆篩選
 6-7周
Primary Cell Bank收集
  • Primary Cell Bank (PCB) 準備
  • PCB穩定性鑒定
 12周
  • 候選克隆穩定性傳代培養報告
  • 穩定細胞株測序報告
  • 支原體檢測報告
  • 關鍵材料COA（細胞株構建過程中用到的試劑耗材的COA）
CHO-K1細胞亞授權
  • CHO-K1細胞株報告
  • CHO-K1細胞亞授權協議
 3-5天
  • CHO-K1細胞株文件和檢測報告
  • CHO-K1細胞亞授權協議

案例分享

案例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.

案例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 (Pembrolizumab) 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 (Pembrolizumab) 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 (Pembrolizumab) 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
“百英公司CHOK1BN細胞株是從ECACC獲得貼壁培養的CHO-K1細胞，并取得商業化亞授權權益。按照項目授權，一次授權，一次性授權。”
Jing Cheng
成靜
細胞株構建團隊

FAQs

  • 什么是CHO-K1細胞？

    中國倉鼠卵巢細胞（Chinese hamster ovary，CHO）。自1958年Puke實驗室將此連續傳代細胞進行重新克隆后，建立了我們現在所用的CHO細胞最原始細胞系。隨著時間的沉淀和歷史的選擇，以及對原始CHO細胞系的不同的培養、改造后，現已出現多種生長、表達、代謝以及基因組等具有差異的細胞系。 1

  • 為什么選擇CHO細胞？

    之所以CHO細胞被選擇，除了具有和人類似地翻譯后的修飾，還能夠在無血清及化學限定培養基懸浮培養中以高密度生長；具有產物胞外分泌功能，并且很少分泌自身的內源蛋白，便于下游產物分離純化，并且在延長的發酵周期中維持高水平的蛋白質表達。 2

  • 為什么CHO細胞常用于治療抗體藥物生產？

    現階段用于生產治療性抗體（Abs）的宿主細胞往往是哺乳動物細胞，主要包括：Sp2/0 骨髓瘤細胞、NS0 小鼠骨髓瘤細胞、HEK293人胚胎腎細胞和中國倉鼠卵巢細胞（Chinese hamster ovary，CHO），其中CHO細胞已然成為需要復雜翻譯后修飾的治療性抗體首選制造細胞，占比超過70%。例如，曲妥珠單抗是一種由CHO細胞表達的治療性抗體，對人表皮生長因子受體2 (HER2)具有特異性。

參考文獻

  • 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

服務和產品

更多服務