相关实验视频
Updated: Aug 12, 2026
11:14
Primer-Free Aptamer Selection Using A Random DNA Library
Published on: July 26, 2010
非SELEX选择的阿普他默
Maxim Berezovski1, Michael Musheev, Andrei Drabovich
1Department of Chemistry, York University, Toronto, Ontario M3J 1P3, Canada.
Journal of the American Chemical Society
|February 2, 2006
概括
这项研究引入了一种新的,快速的aptamer选择方法,没有PCR放大,在短短一个小时内达到高亲和力. 这种非SELEX方法加快了发现速度,并将aptamer应用扩展到无法放大的分子.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 传统上,Aptamer选择使用了通过指数式丰富 (SELEX) 的联体的系统演化.
- SELEX涉及代分区和PCR放大步骤,这些步骤耗时且限制了应用.
研究的目的:
- 开发和验证一种非SELEX的合体选择方法.
- 通过平衡混合物的非平衡毛细体电泳检测 (NECEEM) 来证明这种新方法的效率和速度.
主要方法:
- 在没有干预PCR放大的情况下,利用NECEEM进行分区步骤.
- 采用非SELEX策略,使用重复的NECEEM分区.
- 应用了这种方法来对目标蛋白质选择DNA体.
主要成果:
- 在仅仅三个NECEEM分区步骤中,在DNA库对目标蛋白的亲和力方面取得了超过4个数量级的改进.
- 由此产生的aptamer亲和度超过了三轮基于NECEEM的SELEX的亲和度.
- 在短短1小时内完成了整个选择过程,比传统的SELEX快得多.
结论:
- 引入了一种极其快速,经济和高效的非SELEX 吸收体选择方法.
- 证明了体可能比以前想象的更丰富.
- 从不适合SELEX的小分子库中选择候选药物的新途径.
相关概念视频
01:25Selectins
Cell adhesion is an essential aspect of multicellularity. While stable cell interactions usually occur between cells of the same type, transient cell interactions occur between cells of different tissue types, such as between neutrophils and endothelial cells. Selectins are one class of cell adhesion molecules (CAMs) that bind carbohydrate ligands to form transient cell adhesion. They are rod-like proteins with a long extracellular part of variable length ending with the lectin domain, which...
01:33Enzyme-Linked Immunosorbent Assay
In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.