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Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells.

Herce, Henry D. ; Schumacher, Dominik ; Schneider, Anselm F. L. ; Ludwig, Anne K. ; Mann, Florian A. ; Fillies, Marion ; Kasper, Marc-André ; Reinke, Stefan ; Krause, Eberhard ; Leonhardt, Heinrich ; Cardoso, M. Cristina ; Hackenberger, Christian P. R. (2017)
Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells.
In: Nature chemistry, 9 (8)
Article, Bibliographie

Abstract

Functional antibody delivery in living cells would enable the labelling and manipulation of intracellular antigens, which constitutes a long-thought goal in cell biology and medicine. Here we present a modular strategy to create functional cell-permeable nanobodies capable of targeted labelling and manipulation of intracellular antigens in living cells. The cell-permeable nanobodies are formed by the site-specific attachment of intracellularly stable (or cleavable) cyclic arginine-rich cell-penetrating peptides to camelid-derived single-chain VHH antibody fragments. We used this strategy for the non-endocytic delivery of two recombinant nanobodies into living cells, which enabled the relocalization of the polymerase clamp PCNA (proliferating cell nuclear antigen) and tumour suppressor p53 to the nucleolus, and thereby allowed the detection of protein-protein interactions that involve these two proteins in living cells. Furthermore, cell-permeable nanobodies permitted the co-transport of therapeutically relevant proteins, such as Mecp2, into the cells. This technology constitutes a major step in the labelling, delivery and targeted manipulation of intracellular antigens. Ultimately, this approach opens the door towards immunostaining in living cells and the expansion of immunotherapies to intracellular antigen targets.

Item Type: Article
Erschienen: 2017
Creators: Herce, Henry D. ; Schumacher, Dominik ; Schneider, Anselm F. L. ; Ludwig, Anne K. ; Mann, Florian A. ; Fillies, Marion ; Kasper, Marc-André ; Reinke, Stefan ; Krause, Eberhard ; Leonhardt, Heinrich ; Cardoso, M. Cristina ; Hackenberger, Christian P. R.
Type of entry: Bibliographie
Title: Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells.
Language: English
Date: August 2017
Journal or Publication Title: Nature chemistry
Volume of the journal: 9
Issue Number: 8
Abstract:

Functional antibody delivery in living cells would enable the labelling and manipulation of intracellular antigens, which constitutes a long-thought goal in cell biology and medicine. Here we present a modular strategy to create functional cell-permeable nanobodies capable of targeted labelling and manipulation of intracellular antigens in living cells. The cell-permeable nanobodies are formed by the site-specific attachment of intracellularly stable (or cleavable) cyclic arginine-rich cell-penetrating peptides to camelid-derived single-chain VHH antibody fragments. We used this strategy for the non-endocytic delivery of two recombinant nanobodies into living cells, which enabled the relocalization of the polymerase clamp PCNA (proliferating cell nuclear antigen) and tumour suppressor p53 to the nucleolus, and thereby allowed the detection of protein-protein interactions that involve these two proteins in living cells. Furthermore, cell-permeable nanobodies permitted the co-transport of therapeutically relevant proteins, such as Mecp2, into the cells. This technology constitutes a major step in the labelling, delivery and targeted manipulation of intracellular antigens. Ultimately, this approach opens the door towards immunostaining in living cells and the expansion of immunotherapies to intracellular antigen targets.

Identification Number: pmid:28754949
Divisions: 10 Department of Biology
10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 29 Aug 2017 06:02
Last Modified: 29 Aug 2017 06:02
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