Developments in biological assays by a noncompetitive detection of peptides

肽非竞争性检测生物测定的进展

• 批准号: 20360368
• 负责人: UEDA Hiroshi
• 金额: $ 7.99万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20360368/
• 关键词: 生物・生体工学; 分析科学; 蛋白質

Using an immunoassay that utilizes the antigen-dependent stabilization of antibody variable (Fv) regions (Open-sandwich immunoassay, OS-IA), we attempted to create systems for non-competitive sensitive detection of small molecular peptides, which was not measurable by conventional sandwich assays. Bypassing hybridoma, by utilizing newly developed phage display system for antibody Fab fragments, we succeeded in directly obtaining good antibodies for OS-IA from conjugate-immunized mice with thyroxine and an antigenic peptide from highly pathogenic influenza virus as antigens. Moreover, by OS-selection which is a method for affinity maturation using unconjugated small molecule antigens based on the OS-IA principle, we could improve the sensitivity for osteocalcin peptide by several hundred fold.

使用利用抗体可变（Fv）区的抗原依赖性稳定化的免疫测定（开放夹心免疫测定，OS-IA），我们试图创建用于小分子肽的非竞争性灵敏检测的系统，这是通过常规夹心测定不可测量的。利用新近开发的噬菌体抗体Fab段展示系统，通过杂交瘤的筛选，成功地从以甲状腺素和高致病性流感病毒抗原肽为抗原的结合免疫小鼠中直接获得了抗OS-IA的良好抗体。此外，通过OS选择，这是一种基于OS-IA原理使用未缀合的小分子抗原进行亲和力成熟的方法，我们可以将骨钙素肽的灵敏度提高数百倍。

项目成果

A new gibberellin detection system in living cells based on VH/VL inte raction of antibody

基于抗体VH/VL相互作用的新型活细胞赤霉素检测系统

• 发表时间: 2008
• 期刊: iochem. Biophys. Res. Commun 376
• 作者: Y. Lee;T. Asami;I. Yamaguchi;H. Ueda and Y. Suzuki
• 通讯作者: H. Ueda and Y. Suzuki

Micro OS-ELISA: Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) integrated into microfluidic device

• DOI: 10.1039/b915516c
• 发表时间: 2010-01-01
• 期刊: LAB ON A CHIP
• 影响因子: 6.1
• 作者: Ihara, Masaki;Yoshikawa, Amane;Ueda, Hiroshi
• 通讯作者: Ueda, Hiroshi

Rapid noncompetitive detection of a small biomarker peptide by open-sandwich enzyme linked immunosorbent assay(OS-ELISA)integrated into microfluidic device

通过集成到微流体装置中的开放夹心酶联免疫吸附测定 (OS-ELISA) 快速非竞争性检测小生物标志物肽

• 发表时间: 2010
• 期刊: Lab.Chip 10
• 作者: M.Ihara;A.Yoshikawa;Y.Wu;H.Takahashi;K.Mawatari;K.Shimura;K.Sato;T.Kitamori;H.Ueda
• 通讯作者: H.Ueda

Formation of highly toxic soluble amyloid beta oligomers by the molecular chaperone prefoldin

• DOI: 10.1111/j.1742-4658.2008.06727.x
• 发表时间: 2008-12
• 期刊: The FEBS Journal
• 作者: M. Sakono;T. Zako;H. Ueda;M. Yohda;M. Maeda

抗体融合タンパク質を用いた低分子センサーの創出

使用抗体融合蛋白创建小分子传感器

• 发表时间: 2008
• 作者: R.Tode;M.Matsuoka;M.Anpo;上田宏
• 通讯作者: 上田宏