Abstract
High-throughput ligand discovery and evolution—via genotype-phenotype linkage strategies—empower molecularly targeted therapy, diagnostics, and fundamental science. Maintaining high-quality target antigen in these selections, particularly for membrane targets, is often a technical challenge. Panning yeast-displayed ligand libraries on intact mammalian cells expressing the molecular target has emerged as an effective strategy. Herein we describe the techniques used to select target-binding ligands via this approach including the use of target-negative cells to deplete non-specific binders and avidity reduction to preferentially select high-affinity ligands.
Original language | English (US) |
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Title of host publication | Methods in Molecular Biology |
Publisher | Humana Press Inc. |
Pages | 303-320 |
Number of pages | 18 |
DOIs | |
State | Published - 2020 |
Publication series
Name | Methods in Molecular Biology |
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Volume | 2070 |
ISSN (Print) | 1064-3745 |
ISSN (Electronic) | 1940-6029 |
Bibliographical note
Funding Information:This chapter describes work funded by the American Cancer Society (130418-RSG-17-110-01-TBG to B.J.H.), the National Institutes of Health (R01 EB023339 to B.J.H.), and the California Tobacco-Related Disease Research Grants Program Office of the University of California (28FT-0072 to L.A.S.).
Publisher Copyright:
© 2020, Springer Science+Business Media, LLC, part of Springer Nature.
Keywords
- Avidity
- Cell panning
- Depletion
- Ligand
- Protein engineering
- Specificity
- Yeast surface display