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Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups

[Image: see text] Biosensors and materials for biomedical applications generally require chemical functionalization to bestow their surfaces with desired properties, such as specific molecular recognition and antifouling properties. The use of modified poly(l-lysine) (PLL) polymers with appended oli...

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Autores principales: Movilli, Jacopo, Rozzi, Andrea, Ricciardi, Roberto, Corradini, Roberto, Huskens, Jurriaan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2018
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6302315/
https://www.ncbi.nlm.nih.gov/pubmed/30412384
http://dx.doi.org/10.1021/acs.bioconjchem.8b00733
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author Movilli, Jacopo
Rozzi, Andrea
Ricciardi, Roberto
Corradini, Roberto
Huskens, Jurriaan
author_facet Movilli, Jacopo
Rozzi, Andrea
Ricciardi, Roberto
Corradini, Roberto
Huskens, Jurriaan
author_sort Movilli, Jacopo
collection PubMed
description [Image: see text] Biosensors and materials for biomedical applications generally require chemical functionalization to bestow their surfaces with desired properties, such as specific molecular recognition and antifouling properties. The use of modified poly(l-lysine) (PLL) polymers with appended oligo(ethylene glycol) (OEG) and thiol-reactive maleimide (Mal) moieties (PLL-OEG-Mal) offers control over the presentation of functional groups. These reactive groups can readily be conjugated to, for example, probes for DNA detection. Here we demonstrate the reliable conjugation of thiol-functionalized peptide nucleic acid (PNA) probes onto predeposited layers of PLL-OEG-Mal and the control over their surface density in the preceding synthetic step of the PLL modification with Mal groups. By monitoring the quartz crystal microbalance (QCM) frequency shifts of the binding of complementary DNA versus the density of Mal moieties grafted to the PLL, a linear relationship between probe density and PLL grafting density was found. Cyclic voltammetry experiments using Methylene Blue-functionalized DNA were performed to establish the absolute probe density values at the biosensor surfaces. These data provided a density of 1.2 × 10(12) probes per cm(2) per % of grafted Mal, thus confirming the validity of the density control in the synthetic PLL modification step without the need of further surface characterization.
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spelling pubmed-63023152018-12-25 Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups Movilli, Jacopo Rozzi, Andrea Ricciardi, Roberto Corradini, Roberto Huskens, Jurriaan Bioconjug Chem [Image: see text] Biosensors and materials for biomedical applications generally require chemical functionalization to bestow their surfaces with desired properties, such as specific molecular recognition and antifouling properties. The use of modified poly(l-lysine) (PLL) polymers with appended oligo(ethylene glycol) (OEG) and thiol-reactive maleimide (Mal) moieties (PLL-OEG-Mal) offers control over the presentation of functional groups. These reactive groups can readily be conjugated to, for example, probes for DNA detection. Here we demonstrate the reliable conjugation of thiol-functionalized peptide nucleic acid (PNA) probes onto predeposited layers of PLL-OEG-Mal and the control over their surface density in the preceding synthetic step of the PLL modification with Mal groups. By monitoring the quartz crystal microbalance (QCM) frequency shifts of the binding of complementary DNA versus the density of Mal moieties grafted to the PLL, a linear relationship between probe density and PLL grafting density was found. Cyclic voltammetry experiments using Methylene Blue-functionalized DNA were performed to establish the absolute probe density values at the biosensor surfaces. These data provided a density of 1.2 × 10(12) probes per cm(2) per % of grafted Mal, thus confirming the validity of the density control in the synthetic PLL modification step without the need of further surface characterization. American Chemical Society 2018-11-09 2018-12-19 /pmc/articles/PMC6302315/ /pubmed/30412384 http://dx.doi.org/10.1021/acs.bioconjchem.8b00733 Text en Copyright © 2018 American Chemical Society This is an open access article published under a Creative Commons Non-Commercial No Derivative Works (CC-BY-NC-ND) Attribution License (http://pubs.acs.org/page/policy/authorchoice_ccbyncnd_termsofuse.html) , which permits copying and redistribution of the article, and creation of adaptations, all for non-commercial purposes.
spellingShingle Movilli, Jacopo
Rozzi, Andrea
Ricciardi, Roberto
Corradini, Roberto
Huskens, Jurriaan
Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title_full Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title_fullStr Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title_full_unstemmed Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title_short Control of Probe Density at DNA Biosensor Surfaces Using Poly(l-lysine) with Appended Reactive Groups
title_sort control of probe density at dna biosensor surfaces using poly(l-lysine) with appended reactive groups
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6302315/
https://www.ncbi.nlm.nih.gov/pubmed/30412384
http://dx.doi.org/10.1021/acs.bioconjchem.8b00733
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