Journal
NATURE COMMUNICATIONS
Volume 11, Issue 1, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-020-16920-3
Keywords
-
Categories
Funding
- NIH/NIGMS Interdepartmental Biotechnology Training Program [T32 GM008334]
- Defense Advanced Research Projects Agency (DARPA) [023504-001]
- Calico
Ask authors/readers for more resources
High-diversity genetically-encoded combinatorial libraries (10(8)-10(13) members) are a rich source of peptide-based binding molecules, identified by affinity selection. Synthetic libraries can access broader chemical space, but typically examine only -10(6) compounds by screening. Here we show that in-solution affinity selection can be interfaced with nano-liquid chromatography-tandem mass spectrometry peptide sequencing to identify binders from fully randomized synthetic libraries of 10(8) members-a 100-fold gain in diversity over standard practice. To validate this approach, we show that binders to a monoclonal antibody are identified in proportion to library diversity, as diversity is increased from 10(6) -10(8). These results are then applied to the discovery of p53-like binders to MDM2, and to a family of 3-19 nM-affinity, alpha/beta-peptide-based binders to 14-3-3. An X-ray structure of one of these binders in complex with 14-3-3 sigma is determined, illustrating the role of beta-amino acids in facilitating a key binding contact.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available