Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing

Di Si,1 Guochao Nie,2– 4 Tamiika K Hurst,1 Carol A Fierke,1 Raoul Kopelman1 1Department of Chemistry, University of Michigan, Ann Arbor, MI, USA; 2School of Physics and Telecommunication Engineering, Yulin Normal University, Yulin, People’s Republic of China; 3China-Ukraine Joint Research Center for...

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Autores principales:	Si D, Nie G, Hurst TK, Fierke CA, Kopelman R
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2021
Materias:	paam hydrogel nanogel nanoparticles carbonic anhydrase encapsulation conjugation zn2+. Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/d579dc87cfb444bfaa0a39924bddb86d
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spelling	oai:doaj.org-article:d579dc87cfb444bfaa0a39924bddb86d2021-12-02T17:37:59ZCombining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing1178-2013https://doaj.org/article/d579dc87cfb444bfaa0a39924bddb86d2021-09-01T00:00:00Zhttps://www.dovepress.com/combining-active-carbonic-anhydrase-with-nanogels-enzyme-protection-an-peer-reviewed-fulltext-article-IJNhttps://doaj.org/toc/1178-2013Di Si,1 Guochao Nie,2– 4 Tamiika K Hurst,1 Carol A Fierke,1 Raoul Kopelman1 1Department of Chemistry, University of Michigan, Ann Arbor, MI, USA; 2School of Physics and Telecommunication Engineering, Yulin Normal University, Yulin, People’s Republic of China; 3China-Ukraine Joint Research Center for Nano Carbon Black, Yulin, People’s Republic of China; 4Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology, Yulin, People’s Republic of ChinaCorrespondence: Guochao NieSchool of Physics and Telecommunication Engineering, Yulin Normal University, Yulin, People’s Republic of ChinaEmail bccu518@ylu.edu.cnRaoul KopelmanDepartment of Chemistry, University of Michigan, 930 N University Avenue, Ann Arbor, MI, USAEmail kopelman@umich.eduBackground: Due to its excellent biocompatibility, the polyacrylamide (PAAm) hydrogel has shown great potential for the immobilization of enzymes used in biomedical applications. The major challenge involved is to preserve, during the immobilization process, both the biological activity and the structural integrity of the enzymes. Here we report, for the first time, a proof-of-concept study for embedding active carbonic anhydrase (CA) into polyacrylamide (PAAm) nanogels. By immobilizing CA in these nanogels, we hope to provide important advantages, such as matrix protection of the CA as well as its targeted delivery, and also for potentially using these nanogels as zinc nano-biosensors, both in-vitro and in-vivo.Methods and Results: Two methods are reported here for CA immobilization: encapsulation and surface conjugation. In the encapsulation method, the common process was improved, so as to best preserve the CA, by 1) using a novel biofriendly nonionic surfactant system (Span 80/Tween 80/Brij 30) and 2) using an Al2O3 adsorptive filtration purification procedure. In the surface conjugation method, blank PAAm nanogels were activated by N-hydroxysuccinimide and the CA was cross-linked to the nanogels. The amount of active CA immobilized in the nanoparticles was quantified for both methods. Per 1 g nanogels, the CA encapsulated nanogels contain 11.3 mg active CA, while the CA conjugated nanogels contain 22.5 mg active CA. Also, the CA conjugated nanoparticles successfully measured free Zn2+ levels in solution, with the Zn2+ dissociation constant determined to be 9 pM.Conclusion: This work demonstrates universal methods for immobilizing highly fragile bio-macromolecules inside nanoparticle carriers, while preserving their structural integrity and biological activity. The advantages and limitations are discussed, as well as the potential biomedical applications.Keywords: PAAm hydrogel, nanogels, nanoparticles, carbonic anhydrase, encapsulation, conjugation, Zn2+Si DNie GHurst TKFierke CAKopelman RDove Medical Pressarticlepaam hydrogelnanogelnanoparticlescarbonic anhydraseencapsulationconjugationzn2+.Medicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 16, Pp 6645-6660 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	paam hydrogel nanogel nanoparticles carbonic anhydrase encapsulation conjugation zn2+. Medicine (General) R5-920
spellingShingle	paam hydrogel nanogel nanoparticles carbonic anhydrase encapsulation conjugation zn2+. Medicine (General) R5-920 Si D Nie G Hurst TK Fierke CA Kopelman R Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
description	Di Si,1 Guochao Nie,2– 4 Tamiika K Hurst,1 Carol A Fierke,1 Raoul Kopelman1 1Department of Chemistry, University of Michigan, Ann Arbor, MI, USA; 2School of Physics and Telecommunication Engineering, Yulin Normal University, Yulin, People’s Republic of China; 3China-Ukraine Joint Research Center for Nano Carbon Black, Yulin, People’s Republic of China; 4Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology, Yulin, People’s Republic of ChinaCorrespondence: Guochao NieSchool of Physics and Telecommunication Engineering, Yulin Normal University, Yulin, People’s Republic of ChinaEmail bccu518@ylu.edu.cnRaoul KopelmanDepartment of Chemistry, University of Michigan, 930 N University Avenue, Ann Arbor, MI, USAEmail kopelman@umich.eduBackground: Due to its excellent biocompatibility, the polyacrylamide (PAAm) hydrogel has shown great potential for the immobilization of enzymes used in biomedical applications. The major challenge involved is to preserve, during the immobilization process, both the biological activity and the structural integrity of the enzymes. Here we report, for the first time, a proof-of-concept study for embedding active carbonic anhydrase (CA) into polyacrylamide (PAAm) nanogels. By immobilizing CA in these nanogels, we hope to provide important advantages, such as matrix protection of the CA as well as its targeted delivery, and also for potentially using these nanogels as zinc nano-biosensors, both in-vitro and in-vivo.Methods and Results: Two methods are reported here for CA immobilization: encapsulation and surface conjugation. In the encapsulation method, the common process was improved, so as to best preserve the CA, by 1) using a novel biofriendly nonionic surfactant system (Span 80/Tween 80/Brij 30) and 2) using an Al2O3 adsorptive filtration purification procedure. In the surface conjugation method, blank PAAm nanogels were activated by N-hydroxysuccinimide and the CA was cross-linked to the nanogels. The amount of active CA immobilized in the nanoparticles was quantified for both methods. Per 1 g nanogels, the CA encapsulated nanogels contain 11.3 mg active CA, while the CA conjugated nanogels contain 22.5 mg active CA. Also, the CA conjugated nanoparticles successfully measured free Zn2+ levels in solution, with the Zn2+ dissociation constant determined to be 9 pM.Conclusion: This work demonstrates universal methods for immobilizing highly fragile bio-macromolecules inside nanoparticle carriers, while preserving their structural integrity and biological activity. The advantages and limitations are discussed, as well as the potential biomedical applications.Keywords: PAAm hydrogel, nanogels, nanoparticles, carbonic anhydrase, encapsulation, conjugation, Zn2+
format	article
author	Si D Nie G Hurst TK Fierke CA Kopelman R
author_facet	Si D Nie G Hurst TK Fierke CA Kopelman R
author_sort	Si D
title	Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
title_short	Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
title_full	Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
title_fullStr	Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
title_full_unstemmed	Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
title_sort	combining active carbonic anhydrase with nanogels: enzyme protection and zinc sensing
publisher	Dove Medical Press
publishDate	2021
url	https://doaj.org/article/d579dc87cfb444bfaa0a39924bddb86d
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Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing

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