OXIDATIVE SURFACE FOOTPRINTING FOR ANTI-TUMOR ANTIBODY-ANTIGEN COMPLEXES

抗肿瘤抗体-抗原复合物的氧化表面足迹

• 批准号: 8361809
• 负责人: ROBERT J WOODS
• 金额: $ 0.18万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361809
• 关键词: Animals; Antibodies; Antigen-Antibody Complex; Antigens; Cells; Chemistry; Collaborations; Complex; Computer Simulation; Data; Fab Immunoglobulins; Funding; Gangliosides; Grant; Human; Mass Spectrum Analysis; Molecular; National Center for Research Resources; Peptides; Preparation; Principal Investigator; Production; Research; Research Infrastructure; Resources; Source; Structure; Surface; Tumor Antibodies; Tumor Markers; United States National Institutes of Health; Universities; Work; cost; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is concerned with an antibody called 14F7, which recognizes the ganglioside N-glycolyl GM3. This molecule is very common in animal cells, but not found in human cells except on certain tumors. It is hence an ideal tumor marker. Previous studies of the Krengel group have characterized the crystal structure of the 14F7 Fab and predicted the antigen complex by computer modeling. The aim of this project is to provide experimental data to further characterize the antibody-antigen complex. We will employ oxidative footprinting to generate experimental data that may be used to compare the theoretical structure of the antibody-antigen complex. The project will start with production and purification of 14F7 Fab fragment from purified mAbs. This part of the project will take place at the Department of Chemistry in Oslo. The 14F7 Fab will then be trypsinized and the tryptic peptides characterized by mass spectrometry in collaboration with Dr. Wolfgang Egge-Jacobsen at the Department of Molecular Biosciences. Once these preparations are completed, the work will be continued at the University of Georgia.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 该项目涉及一种名为 14F7 的抗体，该抗体可识别神经节苷脂 N-乙醇酰基 GM3。这种分子在动物细胞中非常常见，但除某些肿瘤外，在人类细胞中未发现。因此它是一种理想的肿瘤标志物。 Krengel小组之前的研究已经表征了14F7 Fab的晶体结构，并通过计算机建模预测了抗原复合物。该项目的目的是提供实验数据以进一步表征抗体-抗原复合物。 我们将采用氧化足迹来生成可用于比较抗体-抗原复合物的理论结构的实验数据。 该项目将从纯化的 mAb 中生产和纯化 14F7 Fab 片段开始。该项目的这一部分将在奥斯陆化学系进行。然后，与分子生物科学系的 Wolfgang Egge-Jacobsen 博士合作，对 14F7 Fab 进行胰蛋白酶处理，并通过质谱法对胰蛋白酶肽进行表征。一旦这些准备工作完成，这项工作将在佐治亚大学继续进行。