The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
27 09 2019
27 09 2019
Historique:
received:
10
01
2019
accepted:
28
08
2019
entrez:
29
9
2019
pubmed:
29
9
2019
medline:
14
1
2020
Statut:
epublish
Résumé
Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG®- or myc-tag. The ALFA-tag forms a small and stable α-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen with low picomolar affinity. It is ideally suited for super-resolution microscopy, immunoprecipitations and Western blotting, and also allows in vivo detection of proteins. We show the crystal structure of the complex that enabled us to design a nanobody mutant (NbALFA
Identifiants
pubmed: 31562305
doi: 10.1038/s41467-019-12301-7
pii: 10.1038/s41467-019-12301-7
pmc: PMC6764986
doi:
Substances chimiques
Epitopes
0
Proteins
0
Recombinant Fusion Proteins
0
Single-Domain Antibodies
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4403Références
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