Screening and Identification of ssDNA Aptamer for Human GP73
As one tumor marker of HCC, Golgi Protein 73 (GP73) is given more promise in the early diagnosis of HCC, and aptamers have been developed to compete with antibodies as biorecognition probes in different detection system. In this study, we utilized GP73 to screen specific ssDNA aptamers by SELEX tech...
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doaj-11377a8820eb4aedb89a2e15985ed4832020-11-24T23:00:41ZengHindawi LimitedBioMed Research International2314-61332314-61412015-01-01201510.1155/2015/610281610281Screening and Identification of ssDNA Aptamer for Human GP73Jingchun Du0Jianming Hong1Chun Xu2Yuanyuan Cai3Bo Xiang4Chengbo Zhou5Xia Xu6Kingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaKingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaKingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaKingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaDepartment of Laboratory Medicine, The First Affiliated Hospital, Guangzhou Medical University, Guangzhou 510120, ChinaKingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaKingmed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou 510182, ChinaAs one tumor marker of HCC, Golgi Protein 73 (GP73) is given more promise in the early diagnosis of HCC, and aptamers have been developed to compete with antibodies as biorecognition probes in different detection system. In this study, we utilized GP73 to screen specific ssDNA aptamers by SELEX technique. First, GP73 proteins were expressed and purified by prokaryotic expression system and Nickle ion affinity chromatography, respectively. At the same time, the immunogenicity of purified GP73 was confirmed by Western blotting. The enriched ssDNA library with high binding capacity for GP73 was obtained after ten rounds of SELEX. Then, thirty ssDNA aptamers were sequenced, in which two ssDNA aptamers with identical DNA sequence were confirmed, based on the alignment results, and designated as A10-2. Furthermore, the specific antibody could block the binding of A10-2 to GP73, and the specific binding of A10-2 to GP73 was also supported by the observation that several tumor cell lines exhibited variable expression level of GP73. Significantly, the identified aptamer A10-2 could distinguish normal and cancerous liver tissues. So, our results indicate that the aptamer A10-2 might be developed into one molecular probe to detect HCC from normal liver specimens.http://dx.doi.org/10.1155/2015/610281 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jingchun Du Jianming Hong Chun Xu Yuanyuan Cai Bo Xiang Chengbo Zhou Xia Xu |
spellingShingle |
Jingchun Du Jianming Hong Chun Xu Yuanyuan Cai Bo Xiang Chengbo Zhou Xia Xu Screening and Identification of ssDNA Aptamer for Human GP73 BioMed Research International |
author_facet |
Jingchun Du Jianming Hong Chun Xu Yuanyuan Cai Bo Xiang Chengbo Zhou Xia Xu |
author_sort |
Jingchun Du |
title |
Screening and Identification of ssDNA Aptamer for Human GP73 |
title_short |
Screening and Identification of ssDNA Aptamer for Human GP73 |
title_full |
Screening and Identification of ssDNA Aptamer for Human GP73 |
title_fullStr |
Screening and Identification of ssDNA Aptamer for Human GP73 |
title_full_unstemmed |
Screening and Identification of ssDNA Aptamer for Human GP73 |
title_sort |
screening and identification of ssdna aptamer for human gp73 |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2015-01-01 |
description |
As one tumor marker of HCC, Golgi Protein 73 (GP73) is given more promise in the early diagnosis of HCC, and aptamers have been developed to compete with antibodies as biorecognition probes in different detection system. In this study, we utilized GP73 to screen specific ssDNA aptamers by SELEX technique. First, GP73 proteins were expressed and purified by prokaryotic expression system and Nickle ion affinity chromatography, respectively. At the same time, the immunogenicity of purified GP73 was confirmed by Western blotting. The enriched ssDNA library with high binding capacity for GP73 was obtained after ten rounds of SELEX. Then, thirty ssDNA aptamers were sequenced, in which two ssDNA aptamers with identical DNA sequence were confirmed, based on the alignment results, and designated as A10-2. Furthermore, the specific antibody could block the binding of A10-2 to GP73, and the specific binding of A10-2 to GP73 was also supported by the observation that several tumor cell lines exhibited variable expression level of GP73. Significantly, the identified aptamer A10-2 could distinguish normal and cancerous liver tissues. So, our results indicate that the aptamer A10-2 might be developed into one molecular probe to detect HCC from normal liver specimens. |
url |
http://dx.doi.org/10.1155/2015/610281 |
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