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A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Published on: February 16, 2014
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SpyDisplay: A versatile phage display selection system using SpyTag/SpyCatcher technology.
Sarah-Jane Kellmann1, Christian Hentrich1, Mateusz Putyrski1
1Bio-Rad AbD Serotec GmbH, Neuried, Germany.
Mabs
|February 21, 2023
Summary
SpyDisplay is a novel phage display method using SpyTag/SpyCatcher ligation for antibody fragment (Fab) display. This robust system enables efficient selection of high-affinity antibodies and facilitates new applications in protein display.
Area of Science:
- Biotechnology
- Molecular Biology
- Immunology
Background:
- Phage display is a key technique for selecting proteins and antibodies from gene libraries.
- Traditional methods rely on genetic fusion of displayed proteins to phage coat proteins.
Purpose of the Study:
- To introduce SpyDisplay, a new phage display method utilizing SpyTag/SpyCatcher protein ligation.
- To demonstrate the functional display and efficient selection of antibody fragments (Fabs) using SpyDisplay.
- To explore novel applications of SpyDisplay beyond traditional phage display.
Main Methods:
- SpyTagged antibody Fabs were ligated to filamentous phages displaying SpyCatcher fused to the pIII coat protein.
- A library of antibody Fabs was cloned and expressed in engineered E. coli.
- Phage panning was employed to isolate specific high-affinity antibody clones.
Main Results:
- Functional and covalent display of SpyTagged Fabs on phage was confirmed.
- High-affinity antibody clones were rapidly isolated using SpyDisplay and phage panning.
- SpyTagged Fabs proved compatible with modular antibody assembly and direct assay testing.
Conclusions:
- SpyDisplay offers a robust and versatile platform for antibody selection and protein display.
- The method streamlines traditional phage display applications and enables new ones, including N-terminal display and display of TAT-exported proteins.
