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

التفاصيل البيبلوغرافية
العنوان: Combining Active Carbonic Anhydrase with Nanogels: Enzyme Protection and Zinc Sensing
المؤلفون: Raoul Kopelman, Di Si, Tamiika K. Hurst, Guochao Nie, Carol A. Fierke
المصدر: International Journal of Nanomedicine
بيانات النشر: Dove, 2021.
سنة النشر: 2021
مصطلحات موضوعية: Biocompatibility, Polyacrylamide, carbonic anhydrase, Biophysics, Pharmaceutical Science, chemistry.chemical_element, Nanoparticle, Bioengineering, Zinc, Conjugated system, Biomaterials, chemistry.chemical_compound, nanogels, International Journal of Nanomedicine, Carbonic anhydrase, Drug Discovery, Original Research, Carbonic Anhydrases, biology, PAAm hydrogel, Chemistry, Organic Chemistry, Biological activity, General Medicine, Zn2+, Enzymes, Immobilized, Dissociation constant, Chemical engineering, biology.protein, encapsulation, Nanoparticles, conjugation
الوصف: 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+
وصف الملف: text/html
اللغة: English
تدمد: 1178-2013
1176-9114
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::23956cd7c162847ed67b9b3fbc5889ea
http://europepmc.org/articles/PMC8486011
حقوق: OPEN
رقم الأكسشن: edsair.doi.dedup.....23956cd7c162847ed67b9b3fbc5889ea
قاعدة البيانات: OpenAIRE
الوصف
تدمد:11782013
11769114