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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2018
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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. |
format | Online Article Text |
id | pubmed-6302315 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
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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