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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
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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. |
format | Online Article Text |
id | pubmed-9400599 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
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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