酵母を人間化して,複雑な末端シアリ化グリコプロテインを生成する
Stephen R Hamilton1, Robert C Davidson, Natarajan Sethuraman
1GlycoFi Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.
まとめ
エンジニアリングされた酵母 Pichia pastorisは,複雑な,シアライテッドのN-グリカンを持つヒトのグリコタンパク質を生成します. イーストのグリコシル化におけるこの進歩は,エリソポエチンのような機能性再結合タンパク質の生産を可能にします.
科学分野:
- バイオテクノロジー バイオテクノロジー
- グライコバイオロジーは,
- 分子生物学は分子生物学である.
背景:
- Pichia pastorisは,再結合タンパク質生産のための一般的なシステムです.
- ネイティブイーストのグリコシレーションはヒトのグリコシレーションとは異なり,治療用タンパク質の応用を制限しています.
研究 の 目的:
- Pichia pastorisをヒトに似たN-グリカン (N-glycan) の生産のために設計する.
- 分泌されるグリコプロテインにおける高レベルの末端シアリレーションを達成するために.
主な方法:
- 4つの内生酵母グリコシル化遺伝子のノックアウト.
- 人間のN-グリコシル化経路を模倣する14の異質遺伝子の導入.
- 遺伝子組み換え酵母菌株を栽培して,グリコタンパク質を生産する.
主要な成果:
- 酵母特異のグリコシライゼーションを成功裏に除去しました.
- エンジニアリングされた細胞系は,複雑なグリコタンパク質の端末シアリレーションを>90%達成しました.
- 機能的な再結合エリトロポエチンを生成することによって有用性が実証されています.
結論:
- 設計されたPichia pastorisシステムは,ヒトのN-グリコシライゼーションを効果的に複製しています.
- このプラットフォームは,治療用途の複雑なシアライラドグリコプロテインの生産に適しています.
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