Cargando…

CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2

Currently, the development of effective diagnostic reagents as well as treatments against Hepatitis C virus (HCV) remains a high priority. In this study, we have described the development of an alive cell surface -Systematic Evolution of Ligands by Exponential Enrichment (CS-SELEX) technique and scr...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Fang, Hu, Yilan, Li, Dongqing, Chen, Haidan, Zhang, Xiao-Lian
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2780912/
https://www.ncbi.nlm.nih.gov/pubmed/19997645
http://dx.doi.org/10.1371/journal.pone.0008142
_version_ 1782174539448320000
author Chen, Fang
Hu, Yilan
Li, Dongqing
Chen, Haidan
Zhang, Xiao-Lian
author_facet Chen, Fang
Hu, Yilan
Li, Dongqing
Chen, Haidan
Zhang, Xiao-Lian
author_sort Chen, Fang
collection PubMed
description Currently, the development of effective diagnostic reagents as well as treatments against Hepatitis C virus (HCV) remains a high priority. In this study, we have described the development of an alive cell surface -Systematic Evolution of Ligands by Exponential Enrichment (CS-SELEX) technique and screened the functional ssDNA aptamers that specifically bound to HCV envelope surface glycoprotein E2. Through 13 rounds of selection, the CS-SELEX generated high-affinity ssDNA aptamers, and the selected ssDNA aptamer ZE2 demonstrated the highest specificity and affinity to E2-positive cells. HCV particles could be specifically captured and diagnosed using the aptamer ZE2. A good correlation was observed in HCV patients between HCV E2 antigen-aptamer assay and assays for HCV RNA quantities or HCV antibody detection. Moreover, the selected aptamers, especially ZE2, could competitively inhibit E2 protein binding to CD81, an important HCV receptor, and significantly block HCV cell culture (HCVcc) infection of human hepatocytes (Huh7.5.1) in vitro. Our data demonstrate that the newly selected ssDNA aptamers, especially aptamer ZE2, hold great promise for developing new molecular probes, as an early diagnostic reagent for HCV surface antigen, or a therapeutic drug specifically for HCV.
format Text
id pubmed-2780912
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-27809122009-12-08 CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2 Chen, Fang Hu, Yilan Li, Dongqing Chen, Haidan Zhang, Xiao-Lian PLoS One Research Article Currently, the development of effective diagnostic reagents as well as treatments against Hepatitis C virus (HCV) remains a high priority. In this study, we have described the development of an alive cell surface -Systematic Evolution of Ligands by Exponential Enrichment (CS-SELEX) technique and screened the functional ssDNA aptamers that specifically bound to HCV envelope surface glycoprotein E2. Through 13 rounds of selection, the CS-SELEX generated high-affinity ssDNA aptamers, and the selected ssDNA aptamer ZE2 demonstrated the highest specificity and affinity to E2-positive cells. HCV particles could be specifically captured and diagnosed using the aptamer ZE2. A good correlation was observed in HCV patients between HCV E2 antigen-aptamer assay and assays for HCV RNA quantities or HCV antibody detection. Moreover, the selected aptamers, especially ZE2, could competitively inhibit E2 protein binding to CD81, an important HCV receptor, and significantly block HCV cell culture (HCVcc) infection of human hepatocytes (Huh7.5.1) in vitro. Our data demonstrate that the newly selected ssDNA aptamers, especially aptamer ZE2, hold great promise for developing new molecular probes, as an early diagnostic reagent for HCV surface antigen, or a therapeutic drug specifically for HCV. Public Library of Science 2009-12-03 /pmc/articles/PMC2780912/ /pubmed/19997645 http://dx.doi.org/10.1371/journal.pone.0008142 Text en Chen et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chen, Fang
Hu, Yilan
Li, Dongqing
Chen, Haidan
Zhang, Xiao-Lian
CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title_full CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title_fullStr CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title_full_unstemmed CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title_short CS-SELEX Generates High-Affinity ssDNA Aptamers as Molecular Probes for Hepatitis C Virus Envelope Glycoprotein E2
title_sort cs-selex generates high-affinity ssdna aptamers as molecular probes for hepatitis c virus envelope glycoprotein e2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2780912/
https://www.ncbi.nlm.nih.gov/pubmed/19997645
http://dx.doi.org/10.1371/journal.pone.0008142
work_keys_str_mv AT chenfang csselexgenerateshighaffinityssdnaaptamersasmolecularprobesforhepatitiscvirusenvelopeglycoproteine2
AT huyilan csselexgenerateshighaffinityssdnaaptamersasmolecularprobesforhepatitiscvirusenvelopeglycoproteine2
AT lidongqing csselexgenerateshighaffinityssdnaaptamersasmolecularprobesforhepatitiscvirusenvelopeglycoproteine2
AT chenhaidan csselexgenerateshighaffinityssdnaaptamersasmolecularprobesforhepatitiscvirusenvelopeglycoproteine2
AT zhangxiaolian csselexgenerateshighaffinityssdnaaptamersasmolecularprobesforhepatitiscvirusenvelopeglycoproteine2