Analytical Sciences

Strategies for Making Multimeric and Polymeric Bifunctional Protein Conjugates and Their Applications as Bioanalytical Tools

Dani PERMANA,*,** Kosuke MINAMIHATA,* Masahiro GOTO,*,*** and Noriho KAMIYA*,***

*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
***Division of Biotechnology, Center for Future Chemistry, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan

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.

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