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通过组合单域抗体库的分步噬菌体选择创建高度特异性化学诱导蛋白分解系统
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Biochimica
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JoVE Journal Biochimica
Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
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通过组合单域抗体库的分步噬菌体选择创建高度特异性化学诱导蛋白分解系统

DOI:
10.3791/60738-v

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10:17 min

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January 14, 2020

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, , , ,
1Department of Biochemistry and Institute for Protein Design,University of Washington

Capitoli

  • 00:04Titolo
  • 00:47TG1 Cell and Negative Selection Bead Preparation
  • 02:15Positive Selection Bead Preparation
  • 03:33Phage-displayed Nanobody Elution and Input/Output Titrations and Infection
  • 05:15Single Clone Isolation
  • 06:22Anchor Binder Validation
  • 08:13Results: Representative Anchor and Dimerization Binder Characterization
  • 09:39Conclusion

Summary

Traduzione automatica

Traduzione automatica

对于任何给定的小分子配体，创建具有所需亲和力和特异性的化学诱导蛋白二聚系统将具有许多生物传感和驱动应用。在这里，我们描述了一种有效的、可通用的方法，通过逐步选择噬菌体显示的组合单域抗体库来对化学诱导的二聚系统进行新工程。

Tags

Keywords: Protein DimerizationBiosensorPhage DisplaySingle-domain AntibodySmall MoleculeCell BehaviorCell Metabolite MonitoringStreptavidin BeadsBiotinylated LigandNegative And Positive Selection

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