受体型酪氨酸蛋白磷酸酶适体的生成

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL Analytical Chemistry Pub Date : 2023-01-03 DOI:10.1021/acs.analchem.2c03988
Jing Lv, Xiaoxiao Zhen, Dandan Li, Guomiao Liao, Zhenhao Long, Nan Zhang, Xiangjun Liu, Tao Bing* and Dihua Shangguan*, 
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引用次数: 1

摘要

Cell-SELEX是一种强大的工具,可以生成适配体,特异性结合活细胞上的天然分子。在这里,我们报道了一个由cell-SELEX产生的适体ZAJ4a。ZAJ4a的分子靶点被富集的细胞selex池拉下,通过基于细胞培养氨基酸(SILAC)的定量蛋白质组学方法进行稳定同位素标记,鉴定为受体型酪氨酸蛋白磷酸酶F (PTPRF)。ZAJ4a在纳米摩尔范围内对表达PTPRF的癌细胞具有很高的结合亲和力。同时,利用ZAJ4a作为分子探针证实了PTPRF在多种癌症细胞系上的高表达,并利用TCGA和GTEx数据库的基因表达谱交互分析(GEPIA2)证实了PTPRF在多种癌症样本中的高表达。这些结果表明,细胞selex产生的适体在分子水平上具有良好的特异性。这种细胞selex和靶标识别策略在识别细胞表面的生物标志物方面显示出巨大的潜力。
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Generation of an Aptamer Targeting Receptor-Type Tyrosine-Protein Phosphatase F

Cell-SELEX is a powerful tool to generate aptamers that specifically bind the native molecules on living cells. Here, we report an aptamer ZAJ4a generated by cell-SELEX. The molecular target of ZAJ4a was pulled down by the enriched cell-SELEX pool and identified to be the receptor-type tyrosine-protein phosphatase F (PTPRF) through a stable isotope labeling using amino acids in cell culture (SILAC)-based quantitative proteomic method. ZAJ4a showed high binding affinity with nanomolar range to cancer cells expressing PTPRF. Meanwhile, PTPRF was proven to highly express on several cancer cell lines using ZAJ4a as a molecular probe and to highly express in many kinds of cancer samples using gene expression profiling interactive analysis (GEPIA2) from the TCGA and GTEx databases. These results indicate that the aptamer generated by cell-SELEX showed good specificity at the molecular level. This cell-SELEX and target identification strategies show great potential for identifying biomarkers on the cell surface.

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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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