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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

Article DOI: 10.3791/60738-v • 10:17 min • January 14th, 2020
January 14th, 2020 •
*1, *1, *1, 1, 1
1Department of Biochemistry and Institute for Protein Design, University of Washington
* These authors contributed equally

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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.

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Chemically Induced Protein Dimerization Systems Phage Selection Combinatorial Single-domain Antibody Library Biosensors Small Molecules Protein Engineering Ligands Cell Behavior Control Real-time Metabolite Monitoring TG1 Electroporation-competent Cells 2YT Medium Negative Selection Beads Streptavidin-coated Magnetic Beads PBS Buffer Casein Biotin
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