Described herein is an enzyme-mediated approach to bioconjugation at nanoparticle (NP) surfaces. This process is enabled by a new synthetic linker compatible with the covalent attachment of alkyne modified substrates, including dyes, peptides and nucleic acids. The methods described herein specifically allow for the linkage of molecules to a DNA-functionalized nanoparticle surface. Enzymatic ligation of molecules to the terminal hydroxyl group of DNA using T4 DNA ligase is achieved through incorporation of a single monophosphate on the approaching substrate. In contrast to previous strategies, the linkers disclosed herein are compatible with alkyne modified molecules of a variety of sizes and charges indicating that the ligase minimally requires the monophosphate and the incoming hydroxyl for conjugation to be successful.
本文描述了一种酶介导的方法,用于在纳米颗粒(NP)表面进行
生物共轭。这一过程通过一种新的合成连接剂实现,该连接剂与炔改性底物(包括
染料、肽和核酸)的共价结合兼容。本文所描述的方法特别允许将分子连接到DNA功能化的纳米颗粒表面。使用T4 DNA连接酶将分子酶接到DNA的末端羟基基团是通过在接近底物上引入单
磷酸基团实现的。与先前的策略相比,本文披露的连接剂与各种大小和电荷的炔改性分子兼容,表明连接酶最少需要单
磷酸基团和进入的羟基才能成功进行共轭。