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Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
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Biochemistry
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JoVE Journal Biochemistry
Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

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

Chapters

  • 00:04Title
  • 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

Automatic Translation

Automatic Translation

Creating chemically induced protein dimerization systems with desired affinity and specificity for any given small molecule ligand would have many biological sensing and actuation applications. Here, we describe an efficient, generalizable method for de novo engineering of chemically induced dimerization systems via the stepwise selection of a phage-displayed combinatorial single-domain antibody library.

Tags

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

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