Molecular analysis of B cell receptor activation by Fab-bridge

Fab-bridge 对 B 细胞受体激活的分子分析

• 批准号: 17570091
• 负责人: YOKOYAMA Miki
• 金额: $ 1.98万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17570091/
• 关键词: B cell receptor; Crosslinking; Antibody

Aim of this study is to set up an experimental system in which we can able to regulate condition of crosslinking of B cell receptor. For this purpose we tried to create "Fab-bridge". Basic concept of Fab-bridge is to connect two Fab portions of anti-mouse IgM monoclonal antibody using linker with different length. Firstly, we planned to raise monoclonal antibody to Fc-portion of myeloma IgM. Although it is considered to be difficult to prepare Fc portion of IgM by papain digestion, Dombrink-Kurzman and Voss reported the Fc preparation by low-temperature papain digestion (Molecular Immunology, 25, 1309-1320(1988)). However, we could not get Fc of IgM by papain-digestion from our IgM. During generation of the Fab portion, Fc portion was completely degraded. So we raised monoclonal antibody against whole IgM. We succeeded in obtaining four monoclonal antibodies. Currently, we are evaluating the ability of these antibody to stimulate mouse splenic B cells.

本研究的目的是建立一个能够调控B细胞受体交联条件的实验系统。为此，我们尝试创建“Fab-bridge”。Fab桥的基本概念是用不同长度的接头将抗鼠IgM单克隆抗体的两个Fab段连接起来。首先，我们计划制备抗骨髓瘤IgM Fc区的单克隆抗体。虽然认为通过木瓜蛋白酶消化制备IgM的Fc部分是困难的，但Dombrink-Kurzman和Voss报道了通过低温木瓜蛋白酶消化制备Fc（Molecular Immunology，25，1309-1320（1988））。但用木瓜蛋白酶消化IgM不能得到Fc。在Fab部分的产生期间，Fc部分完全降解。因此我们研制了抗全IgM的单克隆抗体。我们成功地获得了四个单克隆抗体。目前，我们正在评估这些抗体刺激小鼠脾B细胞的能力。

项目成果

Gangliosides Potentially Inhibit Extracellular Nucleotide Metabolism

神经节苷脂可能抑制细胞外核苷酸代谢

• 发表时间: 2006
• 期刊: Current Medicinal Chemistry 13(19)
• 作者: Osamu Katsumata;et al.;Miki Hara-Yokoyama
• 通讯作者: Miki Hara-Yokoyama

Syntaxin6 separates from GMla-rich membrane microdomain during granule maturation

Syntaxin6 在颗粒成熟过程中与富含 GMla 的膜微区分离

• 发表时间: 2007
• 期刊: Biochemical and Biophysical Research Communications 357
• 作者: Kashiwadani;H.;Moon;K.H.;Lai;M.J.;Osamu Katsumata
• 通讯作者: Osamu Katsumata

Roles of Membrane Domains in the Signaling Pathway for B Cell Survival

膜结构域在 B 细胞存活信号通路中的作用

• 发表时间: 2006
• 期刊: Sphigolipid Biology (Springer-Verlag)
• 作者: Miki Yokoyama;et al.
• 通讯作者: et al.

Sphigolipid Biology (Springer-Verlag)

鞘脂生物学（施普林格出版社）

• 发表时间: 2006
• 作者: Miki Yokoyama;et al.;Miki Yokoyama;Kaori Ueno-Noto et al.;Miki Yokoyama
• 通讯作者: Miki Yokoyama

Recognition of Tandem Sialic Acid Residues by CD38 : A Theoretical Study

CD38 识别串联唾液酸残基：一项理论研究

• 发表时间: 2006
• 期刊: Journal of Computational Chemistry 27(1)
• 作者: Osamu Katsumata;et al.;Miki Hara-Yokoyama;Kaori Ueno-Noto
• 通讯作者: Kaori Ueno-Noto