| Téléchargement | - Voir le manuscrit accepté : Impedance method for detecting HIV-1 protease and screening for its inhibitors using ferrocene-peptide conjugate/Au nanoparticle/single-walled carbon nanotube modified electrode (PDF, 1.5 Mio)
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| DOI | Trouver le DOI : https://doi.org/10.1021/ac801174r |
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| Auteur | Rechercher : Mahmoud, Khaled A.1; Rechercher : Luong, John H. T.1 |
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| Affiliation | - Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
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| Format | Texte, Article |
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| Sujet | environmental |
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| Résumé | A highly sensitive screening assay based on electrochemical impedance spectroscopy (EIS) has been developed for detecting HIV-1 protease (PR) and subsequent evaluation of its corresponding inhibitors at picomolar levels. The assay format was based on the immobilization of the thiol terminated ferrocene(Fc)-pepstatin conjugate on a singlewalled carbon nanotube/gold nanoparticle (SWCNT/AuNP) modified gold electrode. The alteration of the interfacial properties of electrodes upon HIV-1 PR and Fc-pepstatin conjugate interaction was traced by EIS. On the basis of the charge transfer resistance data obtained and using a mixed kinetic and diffusion model, this procedure was capable of detecting picomolar HIV-1 PR owing to the specific binding of this enzyme to Fc modified pepstatin. A competitive inhibition assay format was then performed using four potent HIV-1 PR inhibitors. The estimated inhibition constant (Ki) attested that lopinavir/ritonavir (Ki) 20 ( 3 pM) and saquinavir (Ki = 57 ± 8 pM) even at 10 pM competed strongly with pepstatin for effective binding to HIV-1 PR. Indinavir (Ki = 630 ± 22 pM) only competed well with pepstatin at a much higher concentration (1 nM). No significant inhibitory effect was observed for the fosamprenavir (Ki = 11 ± 0.5 nM) as expected from this pro-drug. Such results agreed well with the values reported in the literature. This assay format is a definite asset for the expedited development of effective HIV-1 PR inhibitors with low molecular weights. |
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| Date de publication | 2008-09-15 |
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| Dans | |
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| Langue | anglais |
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| Numéro du CNRC | NRCC 49938 |
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| Numéro NPARC | 12455580 |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | 7722cbb3-2761-45b3-974b-e2db696165d3 |
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| Enregistrement créé | 2009-10-26 |
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| Enregistrement modifié | 2020-04-15 |
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