RATIONAL DESIGN TO MODULATE ANTIBODY AFFINITY

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 作为治疗剂的单克隆抗体（mAb）因其特异性和其引发免疫应答、拮抗信号传导途径或作为在疾病部位递送细胞毒性化合物的媒介物的能力而被认可。 虽然这些靶向方法显示出改善的PK/PD特性，但许多mAb识别人源性抗原。在治疗剂量下，这些mAb与正常组织上表达的抗原结合，并可产生不良副作用。为了解决这些不良副作用，我们最近开发了一种方法来调节治疗性mAb的抗原亲和力，并使用肿瘤相关蛋白酶在疾病部位激活mAb。 我们表明，切割的“前抗体”恢复的mAb抗原亲和力。 在此，我们正在寻求使用衍射方法鉴定mAb的Fab框架区域内的新型相互作用。 这些分子的成功鉴定将允许我们目前的设计快速优化，促进临床试验，并可能显着改善这一系列的治疗。

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. Monoclonal antibodies (mAbs) as therapeutic agents are recognized for their specificity, and their ability to elicit an immune response, antagonize signaling pathways, or as vehicles to deliver cytotoxic compounds at disease site. While these targeted approaches show improved PK/PD properties, many mAbs recognize antigens that are of human origin. At therapeutic doses, these mAbs bind to antigen expressed on normal tissue and can produce adverse side effects. To address these adverse side effects, we have recently developed a method to modulate the antigen affinity of therapeutic mAbs and use a tumor-associated protease to activate the mAb at the disease site. We show that cleavage of the 'pro-antibody' restores the mAb antigen affinity. Herein, we are seeking to identify novel interactions within the Fab framework region of the mAbs using diffraction methods. Successful identification of these molecules will permit rapid optimization of our current design, facilitate clinical trials and may significantly improve this line of therapy.