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Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting

Although intracellular biomarkers can be imaged with fluorescent dye(s)-labeled antibodies, the use of such probes for precise imaging of intracellular biomarkers in living cells remains challenging due to background noise from unbound probes. Herein, we describe the development of a conditionally a...

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Autores principales: Dai, Yancen, Sato, Yuko, Zhu, Bo, Kitaguchi, Tetsuya, Kimura, Hiroshi, Ghadessy, Farid J., Ueda, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9400599/
https://www.ncbi.nlm.nih.gov/pubmed/36091915
http://dx.doi.org/10.1039/d2sc02355e
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author Dai, Yancen
Sato, Yuko
Zhu, Bo
Kitaguchi, Tetsuya
Kimura, Hiroshi
Ghadessy, Farid J.
Ueda, Hiroshi
author_facet Dai, Yancen
Sato, Yuko
Zhu, Bo
Kitaguchi, Tetsuya
Kimura, Hiroshi
Ghadessy, Farid J.
Ueda, Hiroshi
author_sort Dai, Yancen
collection PubMed
description Although intracellular biomarkers can be imaged with fluorescent dye(s)-labeled antibodies, the use of such probes for precise imaging of intracellular biomarkers in living cells remains challenging due to background noise from unbound probes. Herein, we describe the development of a conditionally active Fab-type Quenchbody (Q-body) probe derived from a monoclonal antibody (DO-1) with the ability to both target and spatiotemporally visualize intracellular p53 in living cells with low background signal. p53 is a key tumor suppressor and validated biomarker for cancer diagnostics and therapeutics. The Q-body displayed up to 27-fold p53 level-dependent fluorescence enhancement in vitro with a limit of detection of 0.72 nM. In fixed and live cells, 8.3- and 8.4-fold enhancement was respectively observed. Furthermore, we demonstrate live-cell sorting based on p53 expression. This study provides the first evidence of the feasibility and applicability of Q-body probes for the live-cell imaging of intrinsically intracellular proteins and opens a novel avenue for research and diagnostic applications on intracellular target-based live-cell sorting.
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spelling pubmed-94005992022-09-08 Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting Dai, Yancen Sato, Yuko Zhu, Bo Kitaguchi, Tetsuya Kimura, Hiroshi Ghadessy, Farid J. Ueda, Hiroshi Chem Sci Chemistry Although intracellular biomarkers can be imaged with fluorescent dye(s)-labeled antibodies, the use of such probes for precise imaging of intracellular biomarkers in living cells remains challenging due to background noise from unbound probes. Herein, we describe the development of a conditionally active Fab-type Quenchbody (Q-body) probe derived from a monoclonal antibody (DO-1) with the ability to both target and spatiotemporally visualize intracellular p53 in living cells with low background signal. p53 is a key tumor suppressor and validated biomarker for cancer diagnostics and therapeutics. The Q-body displayed up to 27-fold p53 level-dependent fluorescence enhancement in vitro with a limit of detection of 0.72 nM. In fixed and live cells, 8.3- and 8.4-fold enhancement was respectively observed. Furthermore, we demonstrate live-cell sorting based on p53 expression. This study provides the first evidence of the feasibility and applicability of Q-body probes for the live-cell imaging of intrinsically intracellular proteins and opens a novel avenue for research and diagnostic applications on intracellular target-based live-cell sorting. The Royal Society of Chemistry 2022-08-01 /pmc/articles/PMC9400599/ /pubmed/36091915 http://dx.doi.org/10.1039/d2sc02355e Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Dai, Yancen
Sato, Yuko
Zhu, Bo
Kitaguchi, Tetsuya
Kimura, Hiroshi
Ghadessy, Farid J.
Ueda, Hiroshi
Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title_full Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title_fullStr Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title_full_unstemmed Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title_short Intra Q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
title_sort intra q-body: an antibody-based fluorogenic probe for intracellular proteins that allows live cell imaging and sorting
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9400599/
https://www.ncbi.nlm.nih.gov/pubmed/36091915
http://dx.doi.org/10.1039/d2sc02355e
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