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Cell Penetration Profiling Using the Chloroalkane Penetration Assay

机译：使用氯烷渗透分析的细胞渗透分析

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摘要

Biotherapeutics are a promising class of molecules in drug discovery, but they are often limited to extracellular targets due to their poor cell penetration. High throughput cell penetration assays are required for the optimization of biotherapeutics for enhanced cell penetration. We developed a HaloTag-based assay called the chloroalkane penetration assay (CAPA), which is quantitative, high throughput, and compartment-specific. We demonstrate the ability of CAPA to profile extent of cytosolic penetration with respect to concentration, presence of serum, temperature, and time. We also used CAPA to investigate structure penetration relationships for bioactive stapled peptides and peptides fused to cell-penetrating sequences. CAPA is not only limited to measuring cytosolic penetration. Using a cell line where HaloTag is localized to the nucleus, we show quantitative measurement of nuclear penetration. Going forward, CAPA will be a valuable method for measuring and optimizing the cell penetration of biotherapeutics.
机译：生物治疗学是药物发现领域中很有前途的一类分子，但由于它们的细胞渗透性差，它们通常限于细胞外靶标。高通量细胞渗透测定是优化生物治疗药物以增强细胞渗透所需的。我们开发了一种基于HaloTag的测定法，称为氯烷烃渗透测定法（CAPA），该测定法定量，高通量且根据隔室而定。我们证明了CAPA能够针对浓度，血清，温度和时间来分析胞质渗透的程度。我们还使用CAPA来研究生物活性固定肽和与细胞穿透序列融合的肽的结构渗透关系。 CAPA不仅限于测量胞质渗透。使用HaloTag定位于细胞核的细胞系，我们显示了核渗透的定量测量。展望未来，CAPA将成为衡量和优化生物治疗药物细胞渗透率的有价值的方法。

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  • 来源
    《Journal of the American Chemical Society》 |2018年第36期|11360-11369|共10页
  • 作者

    Peraro Leila; Deprey Kirsten L.; Moser Matthew K.; Zou Zhongju; Ball Haydn L.; Levine Beth; Kritzer Joshua A.;

  • 作者单位

    Tufts Univ, Dept Chem, Medford, MA 02155 USA;

    Tufts Univ, Dept Chem, Medford, MA 02155 USA;

    Tufts Univ, Dept Chem, Medford, MA 02155 USA;

    Univ Texas Southwestern Med Ctr Dallas, Dept Internal Med, Ctr Autophagy Res, Dallas, TX 75230 USA;

    Univ Texas Southwestern Med Ctr Dallas, Dept Internal Med, Ctr Autophagy Res, Dallas, TX 75230 USA;

    Univ Texas Southwestern Med Ctr Dallas, Dept Internal Med, Ctr Autophagy Res, Dallas, TX 75230 USA;

    Tufts Univ, Dept Chem, Medford, MA 02155 USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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