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Construction of Catalytic-and Electron Transfer-Systems Using Antibody Matrices

使用抗体基质构建催化和电子转移系统

基本信息

批准号：
13650936
负责人：
YAMAGUCHI Hiroyasu
金额：
$ 2.43万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

Monoclonal antibodies for an electron donor-acceptor conjugate were obtained. The hybrid antibody was also prepared by fusing the hybridomas of anti-electron donor (porphyrin) antibody and anti-electron acceptor (quinone) antibody. The hybrid antibody was found to bind both porphyrin and quinone simultaneously, and accelerated the electron transfer from porphyrin to quinone molecule in the antigen-combining site. The complex of the anti-porphyrin antibody with Fe(III)-porphyrin catalyzed oxidation of pyrogallol. The peroxidase activity of this system was found to be high as that of HRP.Monoclonal antibodies for 4,4'-bipyridiniums 1-(carboxypentyl)-1'-methyl-dichloride have been prepared. The complex formation of one of the antibodies with viologen dimer has been studied by a biosensor technique based on surface plasmon resonance (SPR). Enhancement of response signal intensities in SPR is observed by the addition of the antibody solution to the viologen dimer-antibody complex indicating the formation of linear supramolecules between the antibody and viologen dimer. The quantitative amplification of methyl viologen sensing processes is realized by the inhibition of the complex formation between antibodies and viologen dimer by methyl viologen and signal enhancement due to the supramolecular formation of the antibody with viologen dimer. The sensitivity in this system is found to be two orders larger than that in the simple addition of methyl viologen to the antibody immobilized to the surface of the sensor chip.Novel dendrimers have been designed and synthesized by using antibodies. Immunoglobulin M (IgM) which is a pentamer of IgG linked by additional peptides, IgG, and chemically modified IgG were used. Mixing IgM for porphyrin and IgG modified with porphyrin molecule leads to formation of dendritic supramolecules. Antibodies were linked by noncovalent bond with specific molecular recognition of antibodies.

获得了用于电子供体-受体缀合物的单克隆抗体。将抗电子供体（卟啉）抗体和抗电子受体（醌）抗体的杂交瘤融合，制备了杂交抗体。结果表明，该抗体可同时结合卟啉和醌，并在抗原结合位点加速了电子从卟啉向醌分子的转移。抗卟啉抗体与Fe（III）-卟啉的复合物催化焦性没食子酸的氧化。本文还制备了4，4 ′-联吡啶鎓 1-（羧基戊基）-1 ′-甲基二氯化物的单克隆抗体。用表面等离子体共振（SPR）生物传感器技术研究了其中一种抗体与紫精二聚体的复合物形成。通过将抗体溶液添加到紫精二聚体-抗体复合物中观察到SPR中响应信号强度的增强，表明抗体和紫精二聚体之间形成线性超分子。通过甲基紫精抑制抗体和紫精二聚体之间的复合物形成以及由于抗体与紫精二聚体形成超分子而导致的信号增强来实现甲基紫精传感过程的定量放大。该系统的灵敏度比简单地将甲基紫精加入到固定在传感器芯片表面的抗体中的灵敏度高两个数量级。使用免疫球蛋白M（IgM），其是通过另外的肽、IgG和化学修饰的IgG连接的IgG的五聚体。将卟啉修饰的IgM和卟啉修饰的IgG混合，导致树枝状超分子的形成。抗体通过非共价键连接，具有抗体的特异性分子识别。