Raddaoui Nada, Stazzoni Samuele, Möckl Leonhard, Viverge Bastien, Geiger Florian, Engelke Hanna, Bräuchle Christoph, Carell Thomas
Center for Integrated Protein Science (CiPSM), Department of Chemistry, LMU München, Butenandtstrasse 5-13, 81377, München, Germany.
Chembiochem. 2017 Sep 5;18(17):1716-1720. doi: 10.1002/cbic.201700209. Epub 2017 Jul 24.
The in vivo incorporation of alkyne-modified bases into the genome of cells is today the basis for the efficient detection of cell proliferation. Cells are grown in the presence of ethinyl-dU (EdU), fixed and permeabilised. The incorporated alkynes are then efficiently detected by using azide-containing fluorophores and the Cu -catalysed alkyne-azide click reaction. For a world in which constant improvement in the sensitivity of a given method is driving diagnostic advancement, we developed azide- and alkyne-modified dendrimers that allow the establishment of sandwich-type detection assays that show significantly improved signal intensities and signal-to-noise ratios far beyond that which is currently possible.
如今,将炔烃修饰的碱基体内掺入细胞基因组是高效检测细胞增殖的基础。细胞在乙炔基-dU(EdU)存在的情况下生长,然后固定并通透化处理。接着,通过使用含叠氮化物的荧光团和铜催化的炔烃-叠氮化物点击反应,可有效检测掺入的炔烃。在一个给定方法的灵敏度不断提高推动诊断进步的世界里,我们开发了叠氮化物和炔烃修饰的树枝状大分子,可建立三明治型检测分析方法,其信号强度和信噪比显著提高,远超目前的水平。