Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity |
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Authors: | Deng-Liang Wang Yan-Ling Song Zhi Zhu Xi-Lan Li Yuan Zou Hai-Tao Yang Jiang-Jie Wang Pei-Sen Yao Ru-Jun Pan Chaoyong James Yang De-Zhi Kang |
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Affiliation: | 1. The First Clinical Medical College of Fujian Medical University, Fuzhou, China;2. Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China;3. State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China |
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Abstract: | Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy. |
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Keywords: | EGFR, epidermal growth factor receptor SELEX, systematic evolution of ligands by exponential enrichment RPTK, receptor protein tyrosine kinase NSCLC, non-small-cell lung carcinoma TKI, tyrosine kinase inhibitors VEGF, vascular endothelial growth factor ATCC, American Type Culture Collection DMEM, Dulbecco&rsquo s modified Eagle&rsquo s medium WB, washing buffer nt, nucleotides |
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