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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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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. |
format | Online Article Text |
id | pubmed-6111544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT kurumidayoichi developmentofanovelqbodyusinganinvivositespecificunnaturalaminoacidincorporationsystem AT hayashinobuhiro developmentofanovelqbodyusinganinvivositespecificunnaturalaminoacidincorporationsystem |