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Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System

A Q-body capable of detecting target molecules in solutions could serve as a simple molecular detection tool. The position of the fluorescent dye in a Q-body affects sensitivity and therefore must be optimized. This report describes the development of Nef Q-bodies that recognize Nef protein, one of...

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Autores principales: Kurumida, Yoichi, Hayashi, Nobuhiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6111544/
https://www.ncbi.nlm.nih.gov/pubmed/30071687
http://dx.doi.org/10.3390/s18082519
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author Kurumida, Yoichi
Hayashi, Nobuhiro
author_facet Kurumida, Yoichi
Hayashi, Nobuhiro
author_sort Kurumida, Yoichi
collection PubMed
description A Q-body capable of detecting target molecules in solutions could serve as a simple molecular detection tool. The position of the fluorescent dye in a Q-body affects sensitivity and therefore must be optimized. This report describes the development of Nef Q-bodies that recognize Nef protein, one of the human immunodeficiency virus (HIV)’s gene products, in which fluorescent dye molecules were placed at various positions using an in vivo unnatural amino acid incorporation system. A maximum change in fluorescence intensity of 2-fold was observed after optimization of the dye position. During the process, some tryptophan residues of the antibody were found to quench the fluorescence. Moreover, analysis of the epitope indicated that some amino acid residues of the antigen located near the epitope affected the fluorescence intensity.
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spelling pubmed-61115442018-08-30 Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System Kurumida, Yoichi Hayashi, Nobuhiro Sensors (Basel) Article A Q-body capable of detecting target molecules in solutions could serve as a simple molecular detection tool. The position of the fluorescent dye in a Q-body affects sensitivity and therefore must be optimized. This report describes the development of Nef Q-bodies that recognize Nef protein, one of the human immunodeficiency virus (HIV)’s gene products, in which fluorescent dye molecules were placed at various positions using an in vivo unnatural amino acid incorporation system. A maximum change in fluorescence intensity of 2-fold was observed after optimization of the dye position. During the process, some tryptophan residues of the antibody were found to quench the fluorescence. Moreover, analysis of the epitope indicated that some amino acid residues of the antigen located near the epitope affected the fluorescence intensity. MDPI 2018-08-01 /pmc/articles/PMC6111544/ /pubmed/30071687 http://dx.doi.org/10.3390/s18082519 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kurumida, Yoichi
Hayashi, Nobuhiro
Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title_full Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title_fullStr Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title_full_unstemmed Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title_short Development of a Novel Q-body Using an In Vivo Site-Specific Unnatural Amino Acid Incorporation System
title_sort development of a novel q-body using an in vivo site-specific unnatural amino acid incorporation system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6111544/
https://www.ncbi.nlm.nih.gov/pubmed/30071687
http://dx.doi.org/10.3390/s18082519
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