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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Apr 13, 2026

Imaging Protein-protein Interactions in vivo

Imaging Protein-protein Interactions in vivo

Published on: October 10, 2010

高通量FRET亲和选技术 (HTFAST) 用于细胞自由表达结合蛋白的表征.

Sepehr Hejazi, Kimia Noroozi, Vito Jurasic

bioRxiv : the preprint server for biology

|February 23, 2026

概括

此摘要是机器生成的。

我们开发了HTFAST,一种用于快速测量像纳米体这样的蛋白质的结合亲和力的新方法. 这种无细胞技术直接在原始溶解物中起作用,加速新结合蛋白的发展.

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Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

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相关实验视频

Last Updated: Apr 13, 2026

Imaging Protein-protein Interactions in vivo

Imaging Protein-protein Interactions in vivo

Published on: October 10, 2010

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Published on: January 7, 2019

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

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科学领域:

生物技术是生物技术.
蛋白质工程是指蛋白质的工程.
生物物理学的生物物理.

背景情况:

无细胞蛋白合成 (CFPS) 能够快速设计高亲和度结合蛋白.
结合亲和力的高通量表征是一个瓶,特别是在CFPS的未净化蛋白质中.
机器学习指导优化,需要有效的绑定验证.

研究的目的:

开发一种高通量,定量方法,用于对无细胞表达蛋白质的结合亲和性选.
为了使结合蛋白在未经净化的情况下直接在原始溶解酸中进行快速表征.
通过简化查,加速下一代结合蛋白的开发.

主要方法:

开发了使用福斯特共振能量转移 (FRET) 的高通量FRET亲和选技术 (HTFAST).
使用光蛋白融合结合剂和染料标记的抗原来实时测量平衡解离常数.
使用SpyTag003-SpyCatcher003系统优化光对和标签参数.

主要成果:

在原始溶解酸中,HTFAST可靠量化了纳米分子结合亲缘关系.
验证了纳米体平台,包括一种 CD4 结合纳米体 (Nb457).
成功对SARS-CoV-2受体结合域sdabs进行了基准测试,对它们的结合强度进行了排名.
证明了两个有约束力的合作伙伴可以直接表达在CFPS.

结论:

HTFAST提供了一种可扩展,定量和无细胞兼容的高通量亲和力查方法.
该方法非常适合设计-构建-测试-学习 (DBTL) 活动.
加速了各种应用的高亲和度结合蛋白的开发.