Song Hong Y, Ngai Mun H, Song Zhen Y, MacAry Paul A, Hobley Jonathan, Lear Martin J
Department of Chemistry, and Medicinal Chemistry Program of the Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore117543.
Org Biomol Chem. 2009 Sep 7;7(17):3400-6. doi: 10.1039/b904060a. Epub 2009 Jul 3.
The cellular tracking, detection and sensing of protein or antibody movement are important aspects to advance our understanding of biomolecular interactions and activity. Antibodies modified with fluorescent dyes are also valuable tools, especially in immunology research. We describe here a proof-of-principle study of a new water-soluble coumarin probe with a maleimide thiol-reacting unit to fluorescently tag biomolecules. Highlights include: (1) a convenient water-based preparation of N-substituted maleimides, (2) a one-pot preparation of activated maleimido-esters, and (3) a bio-conjugation protocol for the selenol-promoted reduction of native disulfide bonds and the 'site-specific' labelling of antibodies with no significant loss of activity.
蛋白质或抗体运动的细胞追踪、检测与传感是增进我们对生物分子相互作用及活性理解的重要方面。用荧光染料修饰的抗体也是有价值的工具，尤其在免疫学研究中。我们在此描述一项原理验证研究，该研究涉及一种带有马来酰亚胺硫醇反应单元的新型水溶性香豆素探针，用于对生物分子进行荧光标记。要点包括：（1）N-取代马来酰亚胺的便捷水相制备；（2）活性马来酰亚胺酯的一锅法制备；（3）一种生物共轭方案，用于硒醇促进天然二硫键的还原以及抗体的“位点特异性”标记，且活性无显著损失。
