Cargando…
Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat
The development of a new materials platform capable of sustaining the functionality of proteinous sensor molecules over an extended period without being affected by biological contaminants in living systems, such as proteases, is highly demanded. In this study, our primary focus was on fabricating n...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10562976/ https://www.ncbi.nlm.nih.gov/pubmed/37822650 http://dx.doi.org/10.1039/d3ra06108f |
_version_ | 1785118247399456768 |
---|---|
author | Kato, Yuna Iwata, Shuichi Nasu, Yusuke Obata, Akiko Nagata, Kenji Campbell, Robert E. Mizuno, Toshihisa |
author_facet | Kato, Yuna Iwata, Shuichi Nasu, Yusuke Obata, Akiko Nagata, Kenji Campbell, Robert E. Mizuno, Toshihisa |
author_sort | Kato, Yuna |
collection | PubMed |
description | The development of a new materials platform capable of sustaining the functionality of proteinous sensor molecules over an extended period without being affected by biological contaminants in living systems, such as proteases, is highly demanded. In this study, our primary focus was on fabricating new core–shell fibremats using unique polymer materials, capable of functionalizing encapsulated sensor proteins while resisting the effects of proteases. The core-fibre parts of core–shell fibremats were made using a newly developed post-crosslinkable water-soluble copolymer, poly(2-hydroxypropyl methacrylamide)-co-poly(diacetone methacrylamide), and the bifunctional crosslinking agent, adipic dihydrazide, while the shell layer of the nanofibers was made of nylon 6. Upon encapsulating the lactate-sensor protein eLACCO1.1 at the core-fibre part, the fibremat exhibited a distinct concentration-dependent fluorescence response, with a dynamic range of fluorescence alteration exceeding 1000% over the lactate concentration range of 0 to 100 mM. The estimated dissociation constant from the titration data was comparable to that estimated in a buffer solution. The response remained stable even after 5 cycles and in the presence of proteases. These results indicates that our core–shell fibremat platform could serve as effective immobilizing substrates for various sensor proteins, facilitating continuous and quantitative monitoring of various low-molecular-weight metabolites and catabolites in a variety of biological samples. |
format | Online Article Text |
id | pubmed-10562976 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-105629762023-10-11 Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat Kato, Yuna Iwata, Shuichi Nasu, Yusuke Obata, Akiko Nagata, Kenji Campbell, Robert E. Mizuno, Toshihisa RSC Adv Chemistry The development of a new materials platform capable of sustaining the functionality of proteinous sensor molecules over an extended period without being affected by biological contaminants in living systems, such as proteases, is highly demanded. In this study, our primary focus was on fabricating new core–shell fibremats using unique polymer materials, capable of functionalizing encapsulated sensor proteins while resisting the effects of proteases. The core-fibre parts of core–shell fibremats were made using a newly developed post-crosslinkable water-soluble copolymer, poly(2-hydroxypropyl methacrylamide)-co-poly(diacetone methacrylamide), and the bifunctional crosslinking agent, adipic dihydrazide, while the shell layer of the nanofibers was made of nylon 6. Upon encapsulating the lactate-sensor protein eLACCO1.1 at the core-fibre part, the fibremat exhibited a distinct concentration-dependent fluorescence response, with a dynamic range of fluorescence alteration exceeding 1000% over the lactate concentration range of 0 to 100 mM. The estimated dissociation constant from the titration data was comparable to that estimated in a buffer solution. The response remained stable even after 5 cycles and in the presence of proteases. These results indicates that our core–shell fibremat platform could serve as effective immobilizing substrates for various sensor proteins, facilitating continuous and quantitative monitoring of various low-molecular-weight metabolites and catabolites in a variety of biological samples. The Royal Society of Chemistry 2023-10-10 /pmc/articles/PMC10562976/ /pubmed/37822650 http://dx.doi.org/10.1039/d3ra06108f Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/ |
spellingShingle | Chemistry Kato, Yuna Iwata, Shuichi Nasu, Yusuke Obata, Akiko Nagata, Kenji Campbell, Robert E. Mizuno, Toshihisa Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title | Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title_full | Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title_fullStr | Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title_full_unstemmed | Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title_short | Construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(HPMA/DAMA)/ADH-nylon 6 core–shell fibremat |
title_sort | construction of the lactate-sensing fibremats by confining sensor fluorescent protein of lactate inside nanofibers of the poly(hpma/dama)/adh-nylon 6 core–shell fibremat |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10562976/ https://www.ncbi.nlm.nih.gov/pubmed/37822650 http://dx.doi.org/10.1039/d3ra06108f |
work_keys_str_mv | AT katoyuna constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT iwatashuichi constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT nasuyusuke constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT obataakiko constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT nagatakenji constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT campbellroberte constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat AT mizunotoshihisa constructionofthelactatesensingfibrematsbyconfiningsensorfluorescentproteinoflactateinsidenanofibersofthepolyhpmadamaadhnylon6coreshellfibremat |