Modification of Fab Fragments by Dibromopyridazinediones Carrying Mono- and Double-Biotin Functionalities

Takahiko Matsushita; Naoto Maruyama; Tetsuo Koyama; Ken Hatano; Koji Matsuoka

doi:10.1021/acsomega.2c04379

Modification of Fab Fragments by Dibromopyridazinediones Carrying Mono- and Double-Biotin Functionalities

ACS Omega. 2022 Sep 14;7(38):34554-34562. doi: 10.1021/acsomega.2c04379. eCollection 2022 Sep 27.

Authors

Takahiko Matsushita^{1

2

3}, Naoto Maruyama¹, Tetsuo Koyama¹, Ken Hatano^{1

2

3}, Koji Matsuoka^{1

2

3}

Affiliations

¹ Area for Molecular Function, Division of Material Science, Graduate School of Science and Engineering, Saitama University, Sakura, Saitama 338-8570, Japan.
² Medical Innovation Research Unit (MiU), Advanced Institute of Innovative Technology (AIIT), Saitama University, Sakura, Saitama 338-8570, Japan.
³ Health Sciences and Technology Research Area, Strategic Research Center, Saitama University, Sakura, Saitama 338-8570, Japan.

Abstract

To verify the potencies of dibromopyridazinediones with mono- and double-biotin groups, the functions as cysteine-selective biotinylation reagents were evaluated through conjugation with a goat anti-mouse IgG Fab fragment as a functional protein model. The starting Fab was reduced with tris(2-carboxyethyl)phosphine to cleave the disulfide bond and then treated with the reagents. These reagents simultaneously introduced biotin groups into the reduced Fab and re-bridged the disulfide moiety. Furthermore, we demonstrated that the biotin-labeled Fabs were reactive to an antigen and streptavidin.