Development of Fab molecule with the stable scaffold for next generation

开发具有下一代稳定支架的 Fab 分子

• 批准号: 23655160
• 负责人: UEDA Tadashi
• 金额: $ 2.41万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23655160/
• 关键词: 抗体工学; 安定化; アミノ酸改変; ヒト型Fab; 蛋白質の劣化; Fab; 蛋白質工学; 抗体医薬品; 蛋白質; アミノ酸変異; デアミデーション

In order to prepare humanized Fab with the stable scaffold for next generation, we tried to stabilize the constant region of humanized Fab. At first, we showed that Asn138 in L chain of the Fab was easy to deamidate to Asp of Asn residues in the Fab. After preparation of the Fab mutants where Asn138 is mutated to Asp or Ala in L chain of the Fab, respectively, we examined their stabilities against guanidine hydrochloride at pH 6.5 and 25℃. As a result, we found that Asp138Fab was more destabilized than the nonmutant Fab while Ala138Fab had a stability similar to the nonmutant Fab. After heating at pH 9, the residual amount of Ala138Fab was higher than that of the nonmutant Fab.

为了制备具有稳定支架的下一代人源化Fab，我们尝试稳定人源化Fab的恒定区。首先，我们发现Fab的L链Asn138很容易降解为Fab中Asn残基的Asp。将Asn138突变为Fab L链上的Asp和Ala，制备Fab突变体后，在pH 6.5和25℃条件下检测了它们对盐酸胍的稳定性。结果，我们发现Asp138Fab比非突变体Fab更不稳定，而Ala138Fab的稳定性与非突变体Fab相似。在pH 9下加热后，Ala138Fab的残留量高于非突变体Fab。

项目成果

バイオベターな医薬品創製を目指したプロテインエンジニアリング

蛋白质工程旨在创造生物更好的药物

• 发表时间: 2013
• 作者: Man Jae Park;Mamoru Fujitsuka;Tetsuro Majima;植田 正
• 通讯作者: 植田 正

バイオ医薬品の創製とプロテインエンジニアリング

生物制药创造和蛋白质工程

• 发表时间: 2012
• 作者: 阿部義人;植田正
• 通讯作者: 植田正