Selection and targeting of EpCAM protein by ssDNA aptamer.
Aptamers are molecules that reveal highly complex and refined molecular recognition properties. These molecules are capable of binding with high affinity and selectivity to targets, ranging from small molecules to whole living cells. Several aptamers have been selected for targeting cellular protein...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2017-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5731996?pdf=render |
| _version_ | 1819079938761818112 |
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| author | Walhan Alshaer Nidaa Ababneh Mamon Hatmal Heba Izmirli Moujab Choukeife Alaa Shraim Nour Sharar Aya Abu-Shiekah Fadwa Odeh Abeer Al Bawab Abdalla Awidi Said Ismail |
| author_facet | Walhan Alshaer Nidaa Ababneh Mamon Hatmal Heba Izmirli Moujab Choukeife Alaa Shraim Nour Sharar Aya Abu-Shiekah Fadwa Odeh Abeer Al Bawab Abdalla Awidi Said Ismail |
| author_sort | Walhan Alshaer |
| collection | DOAJ |
| description | Aptamers are molecules that reveal highly complex and refined molecular recognition properties. These molecules are capable of binding with high affinity and selectivity to targets, ranging from small molecules to whole living cells. Several aptamers have been selected for targeting cellular proteins and they have also used in developing therapeutics and diagnostic strategies. Epithelial cell adhesion molecule (EpCAM) is considered as a cancer stem cell (CSC) biomarker and one of the most promising targets for aptamer selection against CSCs. In this study, we have developed a ssDNA aptamer with high affinity and selectivity of targeting the EpCAM protein extracellular domain. The SELEX technique was applied and the resulted sequences were tested on EpCAM-positive human gastric cancer cell line, KATO III, and the EpCAM-negative mouse embryonic fibroblast, NIH/3T3 cells. Ep1 aptamer was successfully isolated and showed selective binding on EpCAM-positive KATO III cells when compared to EpCAM-negative NIH/3T3 cells, as observed by the flow cytometry and the confocal imaging results. Additionally, the binding of Ep1 to EpCAM protein was assessed using mobility shifting assay and aptamers-protein docking. Furthermore, the binding affinity of Ep1 was measured against EpCAM protein using EpCAM-immobilized on magnetic beads and showed apparent affinity of 118 nM. The results of this study could suggest that Ep1 aptamer can bind specifically to the cellular EpCAM protein, making it an attractive ligand for targeted drug delivery and as an imaging agent for the identification of cancer cells. |
| first_indexed | 2024-12-21T19:36:57Z |
| format | Article |
| id | doaj.art-d9b29f2d96ce4168b98bc1fdf6490e3f |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T19:36:57Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d9b29f2d96ce4168b98bc1fdf6490e3f2022-12-21T18:52:34ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-011212e018955810.1371/journal.pone.0189558Selection and targeting of EpCAM protein by ssDNA aptamer.Walhan AlshaerNidaa AbabnehMamon HatmalHeba IzmirliMoujab ChoukeifeAlaa ShraimNour ShararAya Abu-ShiekahFadwa OdehAbeer Al BawabAbdalla AwidiSaid IsmailAptamers are molecules that reveal highly complex and refined molecular recognition properties. These molecules are capable of binding with high affinity and selectivity to targets, ranging from small molecules to whole living cells. Several aptamers have been selected for targeting cellular proteins and they have also used in developing therapeutics and diagnostic strategies. Epithelial cell adhesion molecule (EpCAM) is considered as a cancer stem cell (CSC) biomarker and one of the most promising targets for aptamer selection against CSCs. In this study, we have developed a ssDNA aptamer with high affinity and selectivity of targeting the EpCAM protein extracellular domain. The SELEX technique was applied and the resulted sequences were tested on EpCAM-positive human gastric cancer cell line, KATO III, and the EpCAM-negative mouse embryonic fibroblast, NIH/3T3 cells. Ep1 aptamer was successfully isolated and showed selective binding on EpCAM-positive KATO III cells when compared to EpCAM-negative NIH/3T3 cells, as observed by the flow cytometry and the confocal imaging results. Additionally, the binding of Ep1 to EpCAM protein was assessed using mobility shifting assay and aptamers-protein docking. Furthermore, the binding affinity of Ep1 was measured against EpCAM protein using EpCAM-immobilized on magnetic beads and showed apparent affinity of 118 nM. The results of this study could suggest that Ep1 aptamer can bind specifically to the cellular EpCAM protein, making it an attractive ligand for targeted drug delivery and as an imaging agent for the identification of cancer cells.http://europepmc.org/articles/PMC5731996?pdf=render |
| spellingShingle | Walhan Alshaer Nidaa Ababneh Mamon Hatmal Heba Izmirli Moujab Choukeife Alaa Shraim Nour Sharar Aya Abu-Shiekah Fadwa Odeh Abeer Al Bawab Abdalla Awidi Said Ismail Selection and targeting of EpCAM protein by ssDNA aptamer. PLoS ONE |
| title | Selection and targeting of EpCAM protein by ssDNA aptamer. |
| title_full | Selection and targeting of EpCAM protein by ssDNA aptamer. |
| title_fullStr | Selection and targeting of EpCAM protein by ssDNA aptamer. |
| title_full_unstemmed | Selection and targeting of EpCAM protein by ssDNA aptamer. |
| title_short | Selection and targeting of EpCAM protein by ssDNA aptamer. |
| title_sort | selection and targeting of epcam protein by ssdna aptamer |
| url | http://europepmc.org/articles/PMC5731996?pdf=render |
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