SNAP dendrimers: multivalent protein display on dendrimer-like DNA for directed evolution

Kaltenbach, Miriam, Stein, Viktor and Hollfelder, Florian (2011) SNAP dendrimers: multivalent protein display on dendrimer-like DNA for directed evolution. ChemBioChem, 12 14: 2208-2216. doi:10.1002/cbic.201100240


Author Kaltenbach, Miriam
Stein, Viktor
Hollfelder, Florian
Title SNAP dendrimers: multivalent protein display on dendrimer-like DNA for directed evolution
Journal name ChemBioChem   Check publisher's open access policy
ISSN 1439-4227
1439-7633
ISBN 978-9-81-461321-7
Publication date 2011-09-19
Year available 2016
Sub-type Article (original research)
DOI 10.1002/cbic.201100240
Open Access Status Not Open Access
Volume 12
Issue 14
Start page 2208
End page 2216
Total pages 9
Place of publication Weinheim, Germany
Publisher Wiley
Language eng
Abstract Display systems connect a protein with the DNA encoding it. Such systems (e.g., phage or ribosome display) have found widespread application in the directed evolution of protein binders and constitute a key element of the biotechnological toolkit. In this proof-of-concept study we describe the construction of a system that allows the display of multiple copies of a protein of interest in order to take advantage of avidity effects during affinity panning. To this end, dendrimer-like DNA is used as a scaffold with docking points that can join the coding DNA with multiple protein copies. Each DNA construct is compartmentalised in water-in-oil emulsion droplets. The corresponding protein is expressed, in vitro, inside the droplets as a SNAP-tag fusion. The covalent bond between DNA and the SNAP-tag is created by reaction with dendrimer-bound benzylguanine (BG). The ability to form dendrimer-like DNA straightforwardly from oligonucleotides bearing BG allowed the comparison of a series of templates differing in size, valency and position of BG. In model selections the most efficient constructs show recoveries of up to 0.86% and up to 400-fold enrichments. The comparison of mono- and multivalent constructs suggests that the avidity effect enhances enrichment by up to fivefold and recovery by up to 25-fold. Our data establish a multivalent format for SNAP-display based on dendrimer-like DNA as the first in vitro display system with defined tailor-made valencies and explore a new application for DNA nanostructures. These data suggest that multivalent SNAP dendrimers have the potential to facilitate the selection of protein binders especially during early rounds of directed evolution, allowing a larger diversity of candidate binders to be recovered.
Keyword Directed evolution
DNA structures
DNA
DNA-protein conjugates
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
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