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.
本文介绍了一种在纳米粒子(NP)表面进行
生物连接的酶介导方法。这种新的合成连接剂可与炔类修饰基质(包括
染料、肽和核酸)共价连接,从而实现这一过程。本文所述的方法特别允许将分子连接到 DNA 功能化的纳米粒子表面。使用 T4 DNA 连接酶将分子与 DNA 的末端羟基进行酶连接,是通过在接近的底物上加入单
磷酸来实现的。与以往的策略不同,本文公开的连接体与各种大小和电荷的炔修饰分子兼容,这表明连接酶只需要单
磷酸和进入的羟基就能成功连接。