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工業技術研究院

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技術名稱: 人源化之抗人類幹細胞表面抗原CD34之單株抗體

技術簡介

鼠源單株抗體mITRI-10對人類幹細胞表面抗原CD34具有非常高的結合能力,為增加其臨床應用價值,遂以抗體互補變異區移植方式人源化此抗體。先以電腦輔助同源模擬方式建構出mITRI-10抗體變異區之模擬結構,決定出支架區域上可能影響抗原結合的重要胺基酸,保留這些胺基酸與互補決定區域後,其餘置換成人類高度保留之抗體序列,以完成mITRI-10抗體人源化。以哺乳類細胞表現後並純化此抗體蛋白,經測試後顯示此人源化hITRI-10抗體具有與原來鼠原抗體相同之結合人類CD34抗原的結合能力,代表人源化hITRI-10單株抗體可取代鼠源抗體並有更高的臨床應用價值。

Abstract

A murine monoclonal antibody, mITRI-10, showing the highly binding affinity for human CD34 was chosen to be humanized by CDR-grafting approach. The critical residues in the framework region (FR) of the variable regions of both the heavy and light chains of mITRI-10 IgG were identified via a molecular modeling of mITRI-10 built by computer-assisted homology modeling. By retaining these critical residues in the FR, a humanized version, hITRI-10, was generated. The humanized hITRI-10 IgG was then stably expressed in a mammalian cell line and the purified antibody maintained the high antigen binding affinity as the parental murine ITRI-10 antibody. These results suggest that it might be a alternative to mouse anti-CD34 antibodies routinely used clinically.

技術規格

抗體已人源化 2.結合於人類幹細胞CD34之結合強度為KD=6.9 nM 3.抗體可於哺乳類細胞中生產 4.抗體蛋白可以protein A管柱純化,其純度可達90%以上

Technical Specification

The anti-human CD34 antibody has been humanization. 2.The KD value of binding to human CD34 is 6.9 nM. 3.The antibody can be produced by mammalian cell. 4.The antibody can be purified by protein A column and the purity can reach to 90%.

技術特色

此人源化抗人類CD34單株抗體對人類造血幹細胞具有高度的結合能力,並且經過抗體人源化後,可降低應用於人體時可能發生之immunogenecity的機率,增加臨床應用的價值。

應用範圍

造血幹細胞的分離純化裝置 2.針對幹細胞顯影劑的開發 3.增加醫療器材的生物相容性,包括心血管支架、人工器官等 4.幹細胞修復血管與心肌受損之抗體藥物

接受技術者具備基礎建議(設備)

哺乳類細胞培養設備 2.蛋白質純化設備

接受技術者具備基礎建議(專業)

1.哺乳類細胞培養技術 2.蛋白質純化技術

技術分類 生物工程

聯絡資訊

聯絡人:邱偉鈞 標靶藥物技術中心

電話:+886-3-5912148 或 Email:weichiun@itri.org.tw

客服專線:+886-800-45-8899

傳真:+886-3-591-0097

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