A single-round selection of selective DNA aptamers for mammalian cells by polymer-enhanced capillary transient isotachophoresis,Analyst

当前位置： X-MOL 学术 › Analyst › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

A single-round selection of selective DNA aptamers for mammalian cells by polymer-enhanced capillary transient isotachophoresis
Analyst ( IF 3.6 ) Pub Date : 2017-08-15 00:00:00 , DOI: 10.1039/c7an00909g
Kazuki Hirose _{1,

2,

3,

4} , Maho Tsuchida _{1,

2,

3,

4} , Hinako Asakura _{1,

2,

3,

4} , Koji Wakui _{4,

5,

6,

7,

8} , Keitaro Yoshimoto _{4,

5,

6,

7,

8} , Keisuke Iida _{1,

2,

3,

4} , Makoto Sato _{4,

9,

10,

11} , Masami Shibukawa _{1,

2,

3,

4} , Masami Suganuma _{1,

2,

3,

4,

12} , Shingo Saito _{1,

2,

3,

4}

Affiliation

A single-round DNA aptamer selection for mammalian cells was successfully achieved for the first time using a capillary electrophoresis (CE)-based methodology called polymer-enhanced capillary transient isotachophoresis (PectI). The PectI separation yielded a single peak for the human lung cancer cell line (PC-9) complexed with DNA aptamer candidates, which was effectively separated from a free randomized DNA library peak, ensuring no contamination from free DNA in the PC-9–DNA aptamer complex fraction. The DNA aptamer candidates obtained after a single-round selection employing counter selection with HL-60 were proven to bind selectively and form kinetically stable complexes with PC-9 cells. Interestingly, most aptamer candidates showed high binding ability (K_d = 70–350 nM) with different extents of binding on the cell surface. These facts proved that a single-round selection for mammalian cells by PectI is feasible to obtain various types of aptamer candidates, which have high-affinity even for non-overexpressed but unique targets on the cell surface in addition to overexpressed targets.

中文翻译：

通过聚合物增强的毛细管瞬时等速电泳单轮选择哺乳动物细胞的选择性DNA适体

使用基于毛细管电泳（CE）的方法（称为聚合物增强的毛细管瞬态等速电泳（PectI）），首次成功实现了针对哺乳动物细胞的单轮DNA适体选择。PectI分离产生了与DNA适体候选物复合的人肺癌细胞系（PC-9）的单个峰，该峰与有效的随机DNA文库峰有效分离，确保PC-9–DNA中的游离DNA不受到污染适体复合物级分。经证明，使用HL-60反选择单轮选择后获得的DNA适体候选物与PC-9细胞选择性结合并形成动力学稳定的复合物。有趣的是，大多数适体候选物显示出高结合能力（K _d= 70–350 nM），并在细胞表面具有不同程度的结合。这些事实证明，通过PectI对哺乳动物细胞进行单轮选择对于获得各种类型的适体候选物是可行的，这些适体候选物即使对于过度表达的靶标也具有高亲和力，甚至对细胞表面上的独特靶标也具有高亲和力。

更新日期：2017-09-06

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11