Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka, 819-0395, Japan.
Research Unit for Clean Technology, Indonesian Institute of Sciences (LIPI), Kampus LIPI Bandung, Jl. Cisitu, Bandung, 40135, Indonesia.
Anal Sci. 2021 Mar 10;37(3):425-429. doi: 10.2116/analsci.20SCR07. Epub 2021 Jan 15.
Enzymes play a central role in the detection of target molecules in biotechnological fields. Most probes used in detection are bifunctional proteins comprising enzymes and binding proteins conjugated by chemical reactions. To create a highly sensitive detection probe, it is essential to increase the enzyme-to-binding protein ratio in the probe. However, if the chemical reactions required to prepare the probe are insufficiently site-specific, the detection probe may lose functionality. Genetic modifications and enzyme-mediated post-translational modifications (PTMs) can ensure the site-specific conjugation of proteins. They are therefore promising strategies for the production of detection probes with high enzyme contents, i.e., polymeric bifunctional proteins. Herein, we review recent advances in the preparation of bifunctional protein conjugates and polymeric bifunctional protein conjugates for detection. We have summarized research on genetically fused proteins and enzymatically prepared polymeric bifunctional proteins, and will discuss the potential use of protein polymers in various detection applications.
酶在生物技术领域中靶分子的检测中起着核心作用。大多数用于检测的探针都是由通过化学反应连接的酶和结合蛋白组成的双功能蛋白。为了创建高灵敏度的检测探针，必须增加探针中酶与结合蛋白的比例。然而，如果制备探针所需的化学反应不够特异性，检测探针可能会失去功能。遗传修饰和酶介导的翻译后修饰（PTMs）可以确保蛋白质的特异性连接。因此，它们是生产具有高酶含量的检测探针（即聚合双功能蛋白）的有前途的策略。本文综述了用于检测的双功能蛋白缀合物和聚合双功能蛋白缀合物的最新进展。我们总结了关于基因融合蛋白和酶法制备的聚合双功能蛋白的研究，并将讨论蛋白质聚合物在各种检测应用中的潜在用途。
