携带单生物素和双生物素功能的二溴哒嗪二酮对Fab片段的修饰
Modification of Fab Fragments by Dibromopyridazinediones Carrying Mono- and Double-Biotin Functionalities.
作者信息
Matsushita Takahiko, Maruyama Naoto, Koyama Tetsuo, Hatano Ken, Matsuoka Koji
机构信息
Area for Molecular Function, Division of Material Science, Graduate School of Science and Engineering, Saitama University, Sakura, Saitama 338-8570, Japan.
Medical Innovation Research Unit (MiU), Advanced Institute of Innovative Technology (AIIT), Saitama University, Sakura, Saitama 338-8570, Japan.
出版信息
ACS Omega. 2022 Sep 14;7(38):34554-34562. doi: 10.1021/acsomega.2c04379. eCollection 2022 Sep 27.
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.
为验证具有单生物素基团和双生物素基团的二溴哒嗪二酮的效能，通过与作为功能蛋白模型的山羊抗小鼠IgG Fab片段偶联，评估其作为半胱氨酸选择性生物素化试剂的功能。起始Fab用三（2-羧乙基）膦还原以裂解二硫键，然后用试剂处理。这些试剂同时将生物素基团引入还原的Fab中，并重新桥连二硫部分。此外，我们证明生物素标记的Fabs对抗抗原和链霉亲和素具有反应性。
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