クロロアルカン浸透測定法による細胞浸透プロファイリング

Leila Peraro¹, Kirsten L Deprey¹, Matthew K Moser¹

¹Department of Chemistry , Tufts University , Medford , Massachusetts 02155 , United States.

Journal of the American Chemical Society

|August 18, 2018

まとめ

新しいアッセイ,クロロアルカン浸透アッセイ (CAPA) は,バイオセラピー細胞浸透の高通量測定を可能にします. この方法は,細胞と核の浸透を高めるための薬物の投与を最適化するのに役立ちます.

科学分野:

生物化学
分子生物学
薬物の発見

背景:

バイオセラピーは治療的可能性を秘めていますが,細胞内への浸透が悪いため,細胞内標的に到達する際には制限があります.
バイオセラピーの細胞浸透性と有効性を最適化するために,高通量アッセイの開発は極めて重要です.

研究の目的:

バイオセラピーの細胞浸透を測定するための定量的な,高通量,コンパートメント固有の測定法を開発し,検証する.
細胞溶液の浸透に影響を与える要因を調査し,ペプチドの構造-浸透関係を調査する.

主な方法:

クロロアルカン浸透測定法 (CAPA) と呼ばれるHaloTagベースの測定法の開発.
様々な条件下 (濃度,血清,温度,時間) で細胞溶液の浸透を定量的にプロファイルするためにCAPAを使用する.
核にハロタグを配置した細胞線を用いて核浸透を評価する

主要な成果:

CAPAは,細胞溶液の浸透率とその実験条件への依存を量化することに成功しました.
ステープルペプチドと細胞に浸透するペプチド融合の構造-浸透関係を調査した.
核浸透の定量的な測定が実証され,アッセイの有用性を拡大しました.

結論:

クロロアルカン浸透測定法 (CAPA) は,バイオセラピーの細胞浸透を測定し,最適化するための多用途かつ効果的なツールです.
CAPAは,浸透と構造-活動関係に影響を与える要因の研究を容易にする.
このアッセイは細胞に浸透するバイオセラピーの開発を進めるのに重要な価値を持っています.

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