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Mating-based Overexpression Library Screening in Yeast
Published on: July 6, 2018
Mining a yeast library for brain endothelial cell-binding antibodies.
Xin Xiang Wang1, Yong Ku Cho, Eric V Shusta
1Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Nature Methods
|January 9, 2007
Summary
Researchers used yeast surface display to find human antibodies targeting living cell membranes. This method identified 34 antibodies that bind to rat brain endothelial cells, with potential for antigen characterization.
Area of Science:
- Biotechnology
- Immunology
- Cell Biology
Background:
- Identifying antibodies that target specific cell surface proteins is crucial for diagnostics and therapeutics.
- Yeast surface display is a powerful platform for antibody discovery and engineering.
Purpose of the Study:
- To utilize yeast surface display for identifying novel antibodies against plasma membranes of living cells.
- To characterize the binding affinity and internalization potential of identified antibodies.
- To develop a new yeast display immunoprecipitation method for antigen identification.
Main Methods:
- Employing yeast surface display with a nonimmune human single-chain antibody library for antibody selection.
- Performing yeast panning against rat brain endothelial cells to isolate specific binders.
- Utilizing a novel yeast display immunoprecipitation technique for initial antigen characterization.
Main Results:
- Successfully identified 34 unique lead antibodies from the human single-chain antibody library.
- Demonstrated that these antibodies bind to rat brain endothelial cells with a dissociation constant (K(d)) of 82 +/- 15 nM.
- Observed that some identified antibodies are internalized into the target cells, suggesting potential for intracellular delivery.
Conclusions:
- Yeast surface display is an effective method for discovering antibodies that recognize cell surface targets on living cells.
- The identified antibodies show promise for applications in cell-based assays, diagnostics, and targeted therapies.
- The developed yeast display immunoprecipitation procedure offers a valuable tool for preliminary antigen identification.
